proteinas

Proteínas: estruturas, funções e aplicações

As proteínas são macromoléculas formadas por conjuntos de aminoácidos. Essas moléculas apresentam uma grande diversidade de funções moleculares em todos os organismos vivos.

As Proteínas

As proteínas são macromoléculas formadas por conjuntos de aminoácidos. Essas moléculas apresentam uma grande diversidade de funções moleculares em todos os organismos vivos.

Essa diversidade é observada graças às forças evolutivas, em que todas as espécies estão submetidas.

Essas macromoléculas são as responsáveis por processos fundamentais da vida de um organismo, e são estudadas há mais de 50 anos.

Durante esse longo tempo de estudo, foi observado que as proteínas são uniformes, e apenas em alguns casos, é possível observar uma variável abrupta, que é decorrente deacidentes evolutivos”, em que a proteína apresenta uma característica muito diferente de seu grupo.

Caso esseacidenteseja benéfico para o organismo, existe uma grande chance dessa proteína se perpetuar, caso não, a chance dessa mutação se perpetuar se torna pequena.

A análise profunda das proteínas é um processo realizado dentro da Biologia Molecular, em que são observados como esses elementos, formados por carbono, hidrogênio, oxigênio e nitrogênio apresentam diferentes estruturas tridimensionais tão bem ajustadas.

As proteínas participam ativamente de: catálise enzimática, transporte e estoque de moléculas, movimento, suporte mecânico, proteção imune, sinalização intra e extracelular, entre outros.

Essas moléculas são altamente reguladas a níveis molecular e celular, assim, a regulação celular das funções da proteína envolve diferentes expressões gênicas, modificações pós-traducionais e cascatas de sinalização (4).

Já as regulações a nível molecular, são realizadas de acordo com o ambiente em que está a proteína, que é ideal para o seu sítio funcional.

O ajuste de cada proteína é devido a codificação das sequências de aminoácidos, o que garante a cada proteína uma função e também estrutura diferenciada.

A predição dessas estruturas é um elemento imprescindível na Biologia Molecular para projetar as estruturas tridimensionais de uma proteína, pois assim é possível compreender como essa proteína age no organismo, como é correlacionada com as famílias de proteínas, predição de resíduos de interação, entre outros.

Estruturas das Proteínas

Por serem moléculas complexas, as proteínas possuem níveis de conformação natural, i.e, a sua forma pode se modificar de acordo com a sua função. Existem quatro níveis estruturais: estrutura primária, secundária, terciária e quaternária.

Essas estruturas são subdivididas para o melhor entendimento, em que, a estrutura primária é composta pela sequência de aminoácidos dessa proteína. Já a estrutura secundária demonstra as propriedades da alfa hélice ou beta hélice, nessa estrutura é possível observar o primeiro nível de enrolamento helicoidal.

Na estrutura terciária, conseguimos observar a proteína em forma 3D, além do dobramento da cadeia polipeptídica sobre si mesma, em que é possível distinguir o conjunto de estruturas e conformações termodinamicamente similares, devido ao seu enovelamento global.

Lastly, na estrutura quaternária é possível observar cadeias polipeptídicas, que se agrupam e formam uma estrutura com cadeias interligadas.

Regulação Celular

Dentro das células, as proteínas são responsáveis pela coordenação de processos biológicos cruciais, por este motivo, é importante que todas as moléculas interajam de forma consistente e precisa para manter a homeostase do organismo.

Os processos biológicos mais importantes das proteínas são: diferenciação/viabilidade celular, desenvolvimento, metabolismo, apoptose e autofagia.

Se algum desses processos não estiver bem regulado, pode haver então um impacto na viabilidade celular, and for this reason, existe todo um processo de regulação celular das funções das proteínas, que podem ser:

Através da expressão gênica, em que os níveis de transcrição são regulados;

Por meio de interações com cofatores, ligantes e/ou metabólitos, que podem gerar mudanças conformacionais que alteram as atividades das proteínas;

– Lastly, as modificações pós-traducionais, em que os resíduos de aminoácidos são modificados, e assim há alterações na afinidade e/ou interação das proteínas ao DNA, ou na capacidade de estabilidade da própria proteína.

Modificações de proteínas

Como já informado anteriormente, as proteínas são moléculas muito importantes para o funcionamento ótimo de um organismo.

Por este motivo, a Biologia Molecular estuda profundamente esse assunto. Atualmente já é possível realizar modificações estruturais em proteínas, com a intenção de modificar/otimizar a sua função, gerar controles para reações, utilizar sua função em outro organismo, entre outros.

Para isso, é necessário que o pesquisador saiba como funciona essa proteína, além de se certificar sobre os seus sítios ativos, predição de aminoácidos, design, entre outros.

Quanto ao design, houve um avanço tecnológico significativo e de banco de dados, em que há a possibilidade de previsão da estrutura de uma proteína.

Por meio de algoritmos de modelagem de proteínas e refinamento de sequenciamentos de última geração, houve um aumento no ritmo de determinação da estrutura experimental.

Fusão de Proteínas

Uma das maiores utilizações de proteínas modificadas é justamente com a fusão de proteínas, este tipo de fusão recombinante é essencial para a construção de proteínas bioativas estáveis.

Com o avanço da tecnologia de recombinação de DNA, as proteínas de fusão tomaram conta dos laboratórios de Biologia Molecular, em que atualmente, já é possível fundir proteínas, domínios ou outras sequências de nucleotídeos a outras, produzindo então proteínas de fusão que possuem diversas funções.

Normalmente essas proteínas fusionadas são utilizadas para: purificação de proteínas, biofármacos, compreensão de estrutura, meia-vida, localização, efeito terapêutico, entre outros.

Outra função importante é a predição da estrutura de uma proteína-alvo. A modelagem estrutural é utilizada normalmente para modelar uma estrutura desconhecida.

A predição de estruturas proteicas, principalmente quanto a estrutura quaternária não é um desafio fácil, mesmo com o grande progresso tecnológico, e com a fusão de proteínas esse desafio se torna maior, pois o tamanho, complexidade e flexibilidade da proteína aumenta, therefore, se faz necessário procurar por serviços especializados que consigam elucidar as interações de proteínas de fusão sintética, para desvendar os mecanismos inter-domínio, domínios de ligação de co-fatores, substratos, entre outros.

Referências

1.      Kuhlman B, Bradley P. Advances in protein structure prediction and design. Nat Rev Mol Cell Biol. novembro de 2019;20(11):681–97.

2.      Chen X, Zaro JL, Shen W-C. Fusion protein linkers: property, design and functionality. Adv Drug Deliv Rev. outubro de 2013;65(10):1357–69.

3.      Huang P-S, Boyken SE, Baker D. The coming of age of de novo protein design. Nature. setembro de 2016;537(7620):320–7.

4.      Mazmanian K, Sargsyan K, Lim C. How the Local Environment of Functional Sites Regulates Protein Function. J Am Chem Soc. junho de 2020;142(22):9861–71.

5.      Yu K, Liu C, Kim B-G, Lee D-Y. Synthetic fusion protein design and applications. Biotechnol Adv. 2015;33(1):155–64.

6.      Berezovsky IN. Towards descriptor of elementary functions for protein design. Curr Opin Struct Biol. outubro de 2019;58:159–65.

7.      Coluzza I. Computational protein design: a review. J Phys Condens Matter. abril de 2017;29(14):143001. 

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controls

Control: the fundamental part of an experiment

designing your experiment

We know how an experiment starts, we need to read several scientific articles, pay close attention to the results presented., look at the controls, error bars, Materials and methods, ask questions in theses that contain more explanations., talk to lab colleagues, confirm that the laboratory has the necessary equipment and materials.

After that, you take your Ata notebook and start designing your experiment, you already know more or less how your model organism will react, and you also already have in mind what result you will get.

Controls

you start your experiment, uses the necessary controls, and at the end of the day (or month lol) you get the result, look at him and think "hey".

At this point you see that the result was not what you expected, then you go back to your Ata notebook and make sure everything you wrote was correct.

After that you do at least two more independent experiments, to look at your result and think "yeah".

For me this is the most fun moment in science: because my model organism behaves so differently from other similar organisms?

This is another story.…

Analysis of results

The important thing is that for you to reach the conclusion that your “jeez” moment is true, you need important controls.

You need to make sure that all components used during your experiment are of quality., that are working, and that they, together with your analysis, will give you the possibility to defend this result.

The controls also give you the possibility to analyze your negative result., and make sure it's really a negative result., and not a poorly planned result.

BioLinker works with several controls related to Molecular Biology. Click on here to know more.

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GFP

GFP: Learn the story of BioMol's most popular protein!

The History of Green Fluorescent Protein (well known as GFP which is the acronym in English of Green Fluorescent Protein) is another one of those fascinating stories of "brilliant" discoveries (sorry for the pun ><) that revolutionized science!

The history of GFP

Green Fluorescent Protein

The GFP protein was first isolated from the jellyfish species Aequorea victoria by Japanese pharmacist Osamu Shimomura who 15 years worked near Nagasaki.

Escaping the atomic bomb explosion that hit that city (and that he reports having heard the plane before the bomb exploded), he decided to attend pharmacy, as the pharmaceutical science department at Nagasaki University was completely destroyed and the campus had to be temporarily relocated to a location close to where Shimomura lived.

Development

In the United States in the 60, he began to devote himself to studying the phenomenon of bioluminescence in living water Aequorea victoria, widely found in the north pacific.

He came to collect approximately 10.000 specimens of this living water to extract the substance behind the bioluminescence! Shimomura and his collaborators began to purify the extracts and found a protein that was then named “aquaporin”.

Although, by purifying this protein, they found that in addition to aquaporin, there was another protein that exhibited a green fluorescence! Sim! Herself! A GFP! Aquaporin present in jellyfish emits a blue light that is captured by the protein and converts it into a green light.! (It's no coincidence that it fluoresces under UV light!).

But the story didn't end there! in the early years 90, North American molecular biologist Douglas Prasher was trying to develop GFP probes to detect specific nucleotide sequences.

The funding he had to do his scientific research was about to expire and when he tried to apply for another funding, he received a response from his reviewer that his research would have no contribution to society. (who has never received such a comment in scientific life? ¯_(Tsu)_/¯).

How GFP began to be used in laboratories

Mass Publication of Prasher came to the attention of Martin Chalfie, who had the ambitious idea of ​​expressing GFP in other bodies, and in which he was successful! (Uhuuuu!).

Chalfie's group cloned and transferred the GFP gene to the nematode Caenorhabditis elegans and the bacteria Escherichia coli! This discovery opened up an endless path of GFP applications!

GFP allowed intracellular structures to be observed without the need to use synthetic dyes or fluorescent antibodies (which may require the use of detergents for cell permeabilization, which can also harm them). 

GFP is an extremely powerful biological marker! When exposed under the proper wavelengths, GFP does not need the addition of any enzyme and substrate to flourish.

It already has all the "fluorescence machinery" built into its protein structure, all of this built with just its amino acid sequence! A GFP solution has a yellowish tint under artificial light (found indoors) but just put it under the sun and the "scientific magic" happens and it starts to emit a bright green light! The GFP molecule has already been, inclusive, engineer to bloom in different colors (yellow, blue, cyano etc.)!

Even the work of the group of Chinese-American biochemist Roger Yonchien Tsien has resulted in a considerable part of what we currently know about the mechanism of GFP function.!

In addition to describing its structure, your group engineer and also improved the GFP molecule.

The contributions of Osamu Shimomura, Martin Chalfie and Roger Yonchien Tsien were later recognized and the three were awarded the prize Nobel Prize in Chemistry em 2008!

But the most important lesson that the history of GFP tells us is to believe more in our work as a powerful scientific contribution instrument.! Every discovery or observation is relevant and should be considered as another added chapter in human scientific history.! 😉

You are in need of GFP in your lab at an affordable price and fast delivery? Click here: https://biolinker.tech/produto/proteina-fluorescente-verde-gfp-produzida-em-e-coli/

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peptides

How to get peptides with the Phage Display technique

Phage Display is an in vitro peptide ligand selection technique from genomic libraries, as explained on here.

What is the phage display:

O phage display is a powerful technique for the identification and selection of peptides., proteins or antibodies with high affinity and specificity for a specific target.

First described in 1985, this technology has influenced work and discoveries made in the fields of cell biology, immunology, pharmacology, in addition to allowing the study of the molecular interactions of a single molecule or cell.

How to get quality peptides:

Recently, a article published in Nature Comunication chamado “A minimalistic cyclic ice-binding peptide from phage display”, published by Stevens et al., a new theory was discussed to create an alternative to the development of synthetic mimetics of Ice-binding proteins (IBP) using the phage display.
PPIs are a diverse class of proteins that help the organism survive in the presence of ice in cold climates.. They have different origins in many organisms., including bacteria, fungi, plants, insects and fish. This mechanism occurs because freezing is lethal to most organisms and the formation of ice crystals can damage cell membranes., which results in cell disruption.

Between as IBPs, one of the most used are antifreeze proteins, that inhibit the formation of large ice grains inside cells that can damage cell organelles or cause cell death.
But the synthesis of peptides that mimic interactions between IBPs and ice, particularly in large-scale productions, are very variable these days.
Due to this, an alternative is to use the phage display to find peptides that can simplify this binding cycle, once phage display usually leads to selection of short peptides that are composed of naturally occurring amino acids., which can increase the synthetic accessibility of new ice-binding peptides.

recent results:

And the work demonstrated an excellent application of the phage display to find successful short IBP mimetic peptides, complementing traditional design approaches, demonstrating a diversity of peptides that can be screened for potential binding to ice crystals.
This can be used as a promising tool in industrial applications such as cryobiology, food storage, agriculture and technology for development and maintenance by lyophilization of proteins and immunobiologicals.

I need help to develop, how do I do?

in case you need, we are here to help you, just click on here that we will get in touch with you. Here your product is made-to-measure!

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Phage Display

Phage Display: how to use to get your target peptide

Phage Display is a technique that needs an expert like us to help you perform the synthesis of target peptides!

What is Phage Display?

One of the most effective Molecular Biology techniques explored at this moment is the Phage Display, where peptide ligands are selected from genomic libraries.

O Phage Display is used by us scientists to carry out in vitro peptide selection, therefore, you need to link your molecules of interest to the surface peptides of a filamentous bacteriophage.

how is it used?

Currently in Molecular Biology laboratories, projects are increasingly robust and with this there is an increase in demand for research and development.

Even with the advancement of Molecular Biology over the years, we know this technique is not simple.

So you need to know a qualified professional to identify false positives / negatives, as well as using the correct controls to get the desired results.

Normally, the bacteriophage library obtained with this process is used for target affinity selection (usually another protein) in positive cycles of affinity.

How does it work

The “Peg” and the “Display”

First you need the bacteriophages, the phages (“phages”), to introduce them (“display”) to a "target" (which can be proteins, small molecules, cells etc) that you want these phages to turn on.

This is the most important part of the Phage Display, and it's also where your secret is: these phages are not all the same.


In the phage solution that is mixed with the target, there is a huge amount of phages slightly different from each other, around the same amount of people on earth: billion!

The main difference between them are small peptides from, for example, near 12 amino acids This peptide chain was designed by scientists to be exposed outside the bacteriophage capsule., thus being able to get in touch with the "world" around you.

Image of the phages with the peptides on the capsid

As a goal of Phage Display is finding a peptide that binds with high affinity to the target, you can be sure that among the billions of possibilities of peptides, there will at least be a possibility that it will have a combination of amino acids with the correct shape and electrical charge to bind perfectly to the target.

in short: if you don't have the right key for the lock you want, just keep trying billions of keys until you find the one you need.

Bacteriophage Virus Guards Information

But how to isolate and know which is the right peptide that will be together with the other billions of possibilities? That's why instead of just mixing billions of peptides with the target, the phages that have these peptides are mixed.

because the phages, besides having the peptides, keep the genetic sequence that generated those peptides, o DNA.

So instead of doing very expensive and complicated analyzes using the tiny amount of that ideal peptide that binds to the target, just take advantage of the fact that the bacteriophage is "specialist" in multiplying.

Then just cultivate it, then extract enough viral DNA and sequence it – technique that is now very accessible and cheap.

Brute Force on Lock

The success of this method happens by "brute force". There is no rational analysis of what might be the best peptide that would bind to the target., but rather a huge amount of trial-and-error.

That is why, among the billions of possibilities, will not bind to the target only that high affinity peptide. Thousands of phages will also bind with peptides that bind “more or less” well., along with the peptide you are trying to filter out from the rest – remembering here that, in solution, you don't have just one “target” unit, but also billions of targets equal to each other, coming into contact with the different possibilities of peptides attached to phages!

To get rid of these unwanted peptides, the target needs to be washed.

Then, unlike unwanted phages, the target needs to be on something that holds it in the solution container.

A simple way to do this is to use a very clean plastic container and adsorb the target to it. – What is, em si, a technology apart.

Image demonstrating before and after washing

after washing, the phages that still remain attached to the target are released.

This is done either by changing the overall electrical charge of the solution. (the "ionic strength" – changing the pH, and/or adding ions, like salt) or adding more targets in solution, making them "compete" for binding the "trapped" phages to the adsorbed targets and thus releasing them into solution.

Artificial Selection of the Fittest (to bind to the target)

But after that you will still have thousands of low-affinity target-binding peptides in solution. – which at least were able not to go to disposal after washing.

That's why the Phage Display requires sequential cycles of phage exposure to target: all these recovered bacteriophages are multiplied together in a bacterial culture, and then extracted and exposed again to the target, repeating all over again, in a new cycle.

When it happens, at the time of exposure of the phages to the target, there will no longer be billions of phages different from each other, but thousands or hundreds of phages different from each other, but by the millions!

It is this step in which the competition of the possibilities of peptides takes place.: exists on targets (remember if: there is a target, but with millions of copies of it in solution) just a place to connect, so the possibility of a peptide that binds with more affinity will win the dispute with other possibilities of lesser affinity, leaving it out to go away with the wash!

at the end of 3 cycles, selection is complete. Starting from billions of possibilities, practically only one type of phage will remain: the one with the peptide sequence of high affinity for the protein.

Reaching a Consensus

It's quite common that in the end you end up with more than one phage type that bonded with high affinity, but analyzing the sequence of their peptides, generally, there is a consensus.

I.e: despite punctual amino acid variations here and there in the peptide chain, there is a great general similarity of possibilities.

A biophysical test estimating the degree of binding affinity between the target and each selected peptide possibility (for, for example, isothermal titration calorimetry, “ITC”) always reveals that all possibilities have high affinity, with little variation between them.

In some cases, there is also the possibility of having more than one group of consensus sequences of the peptides, what can indicate a location (place) independent binding on target – which may or may not interact with the binding dynamics of the other peptide consensus sequence with the other binding site.

I don't know how to perform the test, what can I do?

We know all the challenges to carry out this type of test and for us your satisfaction is very important.!

We at BioLinker are a company 100% national, Founded by Specialists in Molecular Biology, and have this on-demand service for you.

We carry out the laboratory routine and all quality control, because our goal is for you to receive your quality product at a click!

Click on our link and contact us!! We look forward to helping you!

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How to Start a Molecular Biology Lab

Now that you are able to define which product you will produce in the molecular biology laboratory, choose the location where your laboratory will be and obtain the necessary licenses, you need to prepare for the purchase of equipment.

molecular biology laboratory

Get ready to buy the equipment!

Biotechnology labs are expensive, because you need to use sophisticated and high-tech machinery.

For Molecular Biology laboratories, you will mainly need analytical and semi analytical scales, vórtex, shaker, centrifuges, high precision pipettes, stoves, autoclave, magnetic stirrer, flow of biosafety, freezers, refrigerators, ice machines, microwave, water purifiers, sonicadores, termoblocos, etc.

I.e, is a huge list of equipment, beyond glassware, reagents, disposable products and materials to pack your product.

Don't skimp on equipment, this can be expensive!

It is not easy and you need to research a lot! If you don't have a lot of financial resources, you can find some used equipment.

But be careful, it is necessary to observe if the equipment is adequate and calibrated, otherwise you will not be able to synthesize a good quality product.

The equipment purchase process must be associated with the layout of your laboratory.

I.e, it is necessary that you look at the blueprint of your laboratory and analyze in which room a procedure will be performed.

This facilitates the development of your product and also avoids failures in the process.

You will not extract DNA next to the cleaning sink, right?

Always have a Quality Management plan in place

This entire process must be associated with a management plan for quality.

So the employee will already know how the flow of product development of your company takes place, in which room he will find the reagent he needs, the equipment, etc.

So you shorten the development time and avoid flaws in the process, generating more profits for your company.

Look for affordable molecular biology lab suppliers

Furthermore, Molecular Biology laboratories need inputs with a high degree of purity, so you need to properly analyze your suppliers.

We know not Brazil, the largest distributors of materials for the Molecular Biology laboratory are international companies.

But with the high dollar and delay in delivery, you can start researching national companies that supply products with a lot of quality too!

Value national molecular biology!

Over the last few years, many national companies were created, and this is very important for new companies, because this way we develop products of excellent quality and with more competitive values.

Another important detail is that with the purchase in national companies, you can maintain active networking with other companies, and even generate new collaborations.

So stay tuned on how to buy quality products at affordable prices.

Our tip is to buy from suppliers that have proof of quality.

Or carry out a model with Quality Management in which the supplier of the inputs makes an analysis on the conditions of your laboratory, input development flow, quality analysis, etc.

Never buy products that do not have quality analysis, otherwise it will be very difficult to identify at which stage your product is losing quality.

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How to Open a Molecular Biology Laboratory?

Molecular Biology laboratories are increasing in recent years, and this is very important for the growth of research in Brazil. We need to encourage both science at universities and private science!

Molecular Biology Laboratory
Molecular Biology Laboratory

BioLinker trajectory

As we talked about before, BioLinker's trajectory was marked by the pandemic caused by Covid-19 (https://biolinker.tech/biolinker-e-proteinas-alvo-do-covid-19/).

We make our research more flexible and with the help of incentives (https://biolinker.tech/projetos-pipe-fapesp-inovadores-que-inspiram/).

We have managed to establish ourselves as the only Startup in Latin America to market the Cell-free kit (https://biolinker.tech/o-que-e-a-tecnologia-cell-free-utilizada-na-biolinker/).

But what we haven’t told you yet is our trajectory to establish the BioLinker laboratory at Cietec.

How to Open a Molecular Biology Laboratory

To open a Molecular Biology laboratory, you need multiple licenses, and also very robust planning, otherwise you will not be able to carry out the necessary experiments and neither will you commercialize your products.

Leaving the gym and starting a business is not an easy task! You need planning, look for investors, find suitable professionals, buy specific equipment (that have exorbitant values), sincere analysis about your competitors and also have a Marketing plan.

And that with just 5 people!

Strategies

First you need to find out what the biotechnology market lacks. You will hardly find anything totally unheard of, but you need a differentiator, what in the case of BioLinker is cell-free technology.

Then you need a suitable place to open your molecular biology laboratory..

This part is difficult in Brazil, since the incentive to private biotechnology companies is not that great when compared to other countries in the world (but it is improving!).

BioLinker decided to start its activities at Cietec, localized not IPEN-USP, due to the well-established infrastructure.

Licenses and Registrations

After, you need to start taking licenses to be able to develop molecular biology products.

We have the operating permit, CQB biosafety quality certificate, License in Sanitary Surveillance and Cetesb.

The Operating Permit is what will allow your molecular biology laboratory to be regularized, i.e, is issued by the city hall of your city.

This proves that your company is authorized to carry out its activities in a specific location, and that its activities are also in accordance with municipal legislation!

Stay tuned to what you will write in your Application for Business Permit, keep in mind what you design for your company.

So you avoid unnecessary wear and tear with the city in a possible inspection.

The Biosafety Quality Certificate (CQB) (http://ctnbio.mctic.gov.br/consultar-processo-cqb#/cqb/consultar-processo) is an accreditation that CTNBio grants to companies to carry out projects and activities with Genetically Modified Organisms.

I.e, to a Molecular Biology laboratory, is an essential certificate.

The Health Surveillance Record is a very important document for anyone working in the Health and / or Food field, then again, stay tuned to your claims, the areas in which you want to work.

Always stay one step ahead of the records, so you don't miss the opportunity for a new project.

We know the quantity of chemical products and also biological agents are used in Molecular Biology

So don't forget to adapt to Cetesb – Environmental Company of the State of São Paulo.

It is much easier for you to adapt to current regulations before you even start your activities.

So you don't have the impact – that can even bring financial losses – to properly regulate their activities.

Planning

It is also important that you are aware that your Startup will grow, and then you always need to find places where it is easier and more economically feasible to make this new space feasible.

Never forget also the importance of implementing a Quality System, so little by little, while your company grows, you just need to add new procedures.

This helps with the growth of the company and also in the awareness of new employees.

It is much easier for an employee to join a company already knowing that they must follow rules and procedures than you need to stop their operations to train 10 or more employees.

Lastly, keep in mind that the organization, planning and strategy are the main components to create a successful company!

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The BioLinker team is growing. Find out all about them!

We were very happy to see the BioLinker team grow!! Our Startup has a wide range of professionals you need to meet!

Time da BioLinker
Time da BioLinker

A BioLinker

BioLinker is a Startup 100% that uses an innovative cell synthesis technology, chamada Cell-free (Click here to learn more: https://cutt.ly/ecW1zNs). This methodology, although decreasing by approximately 80% of time for protein synthesis, it's not a simple protocol, and it needs to be very well laid out, and for this reason, BioLinker is the only company in Latin America that sells the cell-free kit, thus facilitating the work of several researchers, mainly in Brazil.

The Scientific Director

The founder and founder of BioLinker is our Scientific Director, Mona Oliveira, who has a degree in Veterinary Medicine from the Federal University of Bahia (UFBA) and sandwich Doctorate from the Graduate Program in Biological Sciences (Biochemistry) of the University of São Paulo (USP) and the Nanoscience and Nanotechnologies Program of the International postgraduate School Institute Jozef Stefan (Ljubljana-Slovenia).

Mona is an extremely dynamic and outgoing person.. During the time we spent together I started to see how easy she is networking and always smiling. She is the type of professional who solves problems from a board that was made in the wrong size for the shaker to new directions for the company. Always solving problems and looking for new goals. People, this woman never stops!

The Technical Director

O Phelipe Vitalle is the Technical Director of BioLinker, Graduated in Veterinary Medicine and Specialist in genetic engineering and membrane protein synthesis. He is responsible for the development of innovative technologies in the production of biomolecules of commercial interest. If Phelipe tells you that you can't develop a protein, forgets, why can't you. He's the kind of professional you work with one day and walks out with your cheek aching from laughing. He is a focused professional, who always seeks the best result, is meticulous with the techniques, but you know very well how to adapt them when necessary. Even because we know that when you know the theory in depth, it is possible to create new robust protocols.

The Production Director

if i changed 10 sentences over a year with the Mario Rodriguez, our Production Director, It was a lot. He holds a BA in Chemistry and a Ph.D. in Biotechnology with an emphasis on molecular dynamics and protein analysis.. He is a very qualified professional and has always been able to help me organize the chemical products.. Sometimes it seems like Mario isn't listening to you, but he is! because in a short time, on your computer, he will deliver you a work without defects and done with great care and dedication.

The Executive Director

O Sandy Ravbar, is our Executive Director and also one of the creators of BioLinker. He oversees fundraising areas., accounting, sales and administration. I lost count of how many languages ​​Sandi speaks, but I think they are 7! So the language barrier doesn't exist at BioLinker.

Sandi is a very calm and focused professional. When I was still working in person, he was always looking for new partnerships, and even though he has a degree in Economics, he understands the product developed by BioLinker very well.. As we can see by name, he is not Brazilian, he was born in slovenia, but there's something very Brazilian about it, I think it's your wife, Mona Oliveira… There's no way not to get infected with this Bahian woman!

The Business Development Manager

A Fabiana Igansi is our Business Development Manager! She holds a degree in Biological Science from the Catholic University of Pelotas and a Master's in Plant Improvement from the Federal University of Pelotas (UFPEL), that with his experience in Molecular Biology, can talk to all customers. Fabi is a fully dedicated professional, which is always willing to answer any questions from customers and bridge the gap between BioLinker, the product and the customer. We only met online, but during all this time I could see how she believes in the company and is willing to make the BioLinker team grow even more.

Associate Researchers do Time da BioLinker

A Natalia Marchesan is an excellent professional, holds a degree in Pharmaceutical Sciences, Master's Degree in Biochemical-Pharmaceutical Technology from the University of São Paulo (USP) in the area of ​​biomaterials and holds a PhD in Biochemical-Pharmaceutical Technology in the area of ​​biopolymers as adjuvants for delivery of biological products with an emphasis on nanotechnology and lyophilization processes. What you ask Natalia to lyophilize, you can be sure that she can.

She always shows up in the lab very excited and saying a lot of things and you just think: “people, she loves this job”. And besides loving this job (and take amazing pictures) she still does it with quality. Thanks to the dedication of the BioLinker team and Natalia Marchesan, only two companies worldwide market the lyophilized cell-free kit.

To the Iris Todeschini there is no bad weather, our Associate Researcher holds a degree in Veterinary Medicine from the University of São Paulo (USP) with a Master's Degree in Microbiology from the Institute of Biological Sciences (USP), with an emphasis on the structural and functional study of proteins.

may the light end, you can run out of water and you will see Iris laughing and pipetting. This researcher always seeks excellence in her projects and never accepts an average result. She organizes the laboratory, always arrives before everyone else and leaves after everyone has left. This professional has a lot of focus and determination and is the lifeblood of protein production at BioLinker.

I already said that the Otto Heringer could very well be a successful actor, but he chose to be successful in research, and managed! He holds a Bachelor's degree in Chemistry, and specializes in the development of new biomolecules. Also Otto is that guy who will use an old B.O.D and turn it into a new shaker. He's that kind of researcher that just when you walk into the lab you start laughing. He is a professional with diverse talents and a great aptitude for entrepreneurship.

A Natalia Silva I wouldn't even need to present. She was the one who did most of BioLinker's social media arts. Natalia Silva has a degree in Biochemistry with extensive experience in molecular biology and protein synthesis. She is a volunteer contributor to BioLinker and the best digital marketing publisher! Whenever I talked to Natalia I was very calm, she is a super centered person, Calm down, funny and very kind. In addition, she has always managed to combine her knowledge of Molecular Biology with marketing. She is a great inspiration and a great professional..

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Selection of recombinant protein

Covid-19 target proteins and BioLinker

BioLinker's journey began only at the end of 2019, but with all the urgency brought by the pandemic, we stand out and initiate the development of several Covid-19 target proteins.

How it all began…

The pandemic required scientists to develop Covid-19 target proteins, and that was the beginning of BioLinker's journey. We started this work more than 10 months and we are honored with the evolution and visibility:

Image of fluorescent cells of target proteins
343″We are establishing different ways of expressing S proteins, RBD e gene N. Todas importantes proteínas para o diagnóstico de pacientes.

Após o estabelecimento de expressão de proteínas S, RBD e gene N de Covid-19, iniciamos uma colaboração com o Prof. Dr. Marco Antonio Stephano para fazer parte do seu trabalho de desenvolvimento de uma vacina nasal em forma de spray. A colaboração com o Prof. Marco Antonio é muito gratificante não só no âmbito científico, mas também por nos proporcionar a experiência de trabalhar com esse profissional incrível.

Disponibilizamos para a equipe do Prof. Marco Antonio Stephano Covid-19 target proteins, that were developed with the technology cell-free.

Image containing several cartoons, with people in mask, syringe, medicines, and serological support.
Journal of USP – Spray vaccine with application on the nose

Diversity of Projects

But we don't stop there, we started to develop our own Covid-19 diagnostic test, that gained prominence in the media for being a project 100% national:

https://pesquisaparainovacao.fapesp.br/startup_busca_desenvolver_teste_de_diagnostico_da_covid19_totalmente_nacional/1406

https://www.saopaulo.sp.gov.br/ultimas-noticias/covid-19-startup-busca-desenvolver-teste-de-diagnostico-totalmente-nacional/

https://eurekalert.org/pub_releases/2020-06/fda-bss062620.php

How BioLinker was engaged with several Covid-19 related projects, foi um dos seis primeiros projetos selecionados pelo edital PIPE-FAPESP em parceria com a Finep.

O diagnóstico é um kit padronizado de reação de ELISA, para detectar no soro dos pacientes anticorpos circulantes IgG, que estão presentes na fase mais tardia da doença.

Este teste também foi sendo desenvolvido com a tecnologia cell-free.

Um mês depois, anunciamos a finalização de mais um projeto: antígenos virais do SARS-CoV-2, que já são comercializados pela BioLinker há mais de 6 meses nesse on here.

Imagem contendo três frascos de vidro de antigenos virais das proteínas-alvo de covid-19

A empresa cresceu, e na pesquisa do desenvolvimento de proteínas -alvo do Covid-19, contamos mais uma vez com o auxílio da FAPESP, dessa vez com o auxílio da especialista Dr. Natalia Marchesan Bexiga, uma pesquisadora que liofiliza absolutamente tudo o que você quiser! Dá uma olhadinha em: https://biolinker.tech/#produtos-e-servicos

Image of a plaque 96 wells containing the lyophilized product, that was distributed to form a B of BioLinker. On the right, two eppendorfs containing the lyophilized reagent, below the cell-free kit containing the eppendorfs that are on the ice.
Below the two images, the photo of researcher Dra. Natalia Marchesan - PIPE FAPESP - 2020
On the right the BioLinker logo, PIPE-FAPESP and FAPESP.

Graças a essa parceria, conseguimos iniciar a elaboração de kits cell-free e proteínas-alvo do Covid-19 liofilizadas. Aumentando assim a estabilidade dos reagentes e facilitando o transporte, armazenamento e desenvolvimento de projetos em todo o Brasil.

Repercussão da Mídia

O ano passou rápido, trabalhamos muito, mas no início do ano, tivemos mais uma ótima notícia: o laboratório do Prof. Dr. Frank Crespilho, situado no Instituto de Química de São Carlos da USP, com o qual temos uma colaboração está nas últimas fases do desenvolvimento de um teste rápido de Covid-19:

http://www5.iqsc.usp.br/2021/usp-desenvolve-teste-rapido-de-covid-19-para-viabilizar-aplicacao-em-massa/

O grande diferencial desse dispositivo é o seu valor, além de ser um projeto 100% national, já que as proteínas virais do Covid-19 são disponibilizadas pela BioLinker.

Esse teste ganhou as mídias sociais, e depois os grandes canais da imprensa:

https://agencia.fapesp.br/pesquisadores-desenvolvem-teste-popular-de-covid-19-para-ampliar-acesso-ao-diagnostico/35036/

https://saude.estadao.com.br/noticias/geral,usp-desenvolve-teste-que-reduz-custo-para-detectar-anticorpos-da-covid-19,70003590440

Ainda não terminamos essa história, mas eu estou ansiosa pra contar todos os detalhes pra vocês, no próximo post!

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PCR technique: You know the main types?

The PCR technique gained notoriety with the Corona virus pandemic. Before, when the Google search was performed, só observávamos publicações totalmente direcionadas ao público acadêmico, mas isso mudou.

Imagem com a sequência de evento do PCR, com os dizeres

A técnica de PCR foi desenvolvida por Saiki em 1985 e Mullis em 1987. Mas Mullis foi o responsável pelo desenvolvimento do conceito de primer de PCR, que contém uma sequência específica para amplificar o DNA alvo, e utilizou a DNA polimerase termoestável, que foi obtida da bactéria de fontes termais Thermus aquaticus.

Imagine que o DNA é um livro de receitas. Você tem muito apego a esse livro de receitas, então você prefere fazer uma cópia da receita de bolo de cenoura com cobertura de chocolate, essa cópia é o RNA.

Agora vamos para o PCR. Quando você têm o livro de receitas, você procura a página do bolo pelo índice, certo? O índice nesse caso é o primer.

Agora dá então pra fazer a proteína de forma sintética com um equipamento chamado termociclador, que precisa ser ajustado de acordo com o produto que você vai fazer, até porque você não usa o mesmo tempo e temperatura pra cozinha uma lasanha, um bolo ou um pudim, ?

Agora, os tipos mais comuns de PCR:

PCR convencionalé o método utilizado para multiplicar um trecho específico do DNA, até que a sua concentração seja amplificada e possa ser detectada. Na reação deve conter a DNA polimerase, íons de Mg2+ e desoxinucleotídeos (ATP, TTP, CTP e GTP).

RT-PCRA técnica de transcrição reversa e amplificação é utilizada para avaliar a expressão gênica a partir do mRNA.

Multiplex PCRSão utilizados diversos DNA alvo para uma única reação e diversos primers específicos para cada fragmento gênico escolhido.

Nested PCRTécnica interessante caso você não consiga amplificar apenas um fragmento gênico na sua reação, ou se a amplificação não é suficiente para a detecção.

I.e, é realizado um PCR, e após a reação um novo PCR com primers internos, aumentando assim a especificidade.

PCR isotérmicoAmplificação isotérmica mediada por loop (Loop-mediated isothermal amplificationLAMP), utiliza enzima que permite amplificação isotérmica, apresenta alta especificidade, sensibilidade, rapidez e custo reduzido.

O resultado da amplificação é visualizado no próprio tubo, a olho nu, que é a técnica utilizada no BioTreco e também no Kit de detecção de Covid-19 que a BioLinker é parceira.

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Innovative projects with FAPESP funding that inspire!

Cartoon image of test tubes and volumetric flask.
Below the BioLinker logo, PIPE-FAPESP and Fapesp.

A BioLinker, with the support of FAPESP financing – PIPE captured more than 1 million for the development of its portfolio of products and services. Here we are going to present a little bit about two projects supported by this program!

The incentive to startups and other biotechnology companies is fundamental for the development of new national technologies.

Prokaryon: the revolution in the form of protein purification!

The technology that allows protein purification inside a microchip! During purification special aptamers are used:
1- Immobilisation of aptamers
2- Affinity Purification
3- Protein elution

Image of operation inside the microchip:
1- aptamer immobilization
2- PH affinity purification 6.4
3- Protein elution at pH 4.6

To the right of the image the BioLinker logos, PIPE-FAPESP and FAPESP.

This project is carried out by Dr.. Mona Oliveira, the Scientific Director of BioLinker.
The initial results are promising. The technology involved was able to efficiently purify Immunoglobulin G from aptamers:

3D image of the aptamer associated with the immunoglobulin G protein

Kits cell-free lyophilized!

Lyophilization is a process that allows the storage of reagents for longer and at temperatures more compatible with more accessible logistics.
This project is carried out by Dr.. Natalia Marchesan, the associate researcher at BioLinker.

Image of a plaque 96 wells containing the lyophilized product, that was distributed to form a B of BioLinker. On the right, two eppendorfs containing the lyophilized reagent, below the cell-free kit containing the eppendorfs that are on the ice.
Below the two images, the photo of researcher Dra. Natalia Marchesan - PIPE FAPESP - 2020
On the right the BioLinker logo, PIPE-FAPESP and FAPESP.

Our team of specialists develops freeze-drying protocols and formulations that guarantee the quality of our products associated with logistical accessibility.

Antigen production from Covid-10 FINEP / FAPESP

Our team of researchers is working hard in this quarantine to help fight Covid-19 with FAPESP funding. Viral antigens are produced and lyophilized for use in innovative research that seeks solutions to fight the SARS-CoV-2 pandemic:

1- MasterEasey PCR-LAMP isotérmico: molecular RNA detection kit. Reverse transcriptase MMLV e Bst Polymerase;

2- Antígenos SARS-CoV-2: we produce 6 viral antigens SARS-CoV-2 (RBD, 3C Protease, Methyltransferase, RdR polymerase, N, E e M);

3- Covid-19 ELISA test: detection kits for Immunoglobulin G against SARS-CoV-2.

Image of three tubes containing lyophilized covid-19 antigens that are part of the FAPESP project.

This project is carried out in partnership with Biobreyer and Tovem Biotech, and the main researcher is Dr. Phelipe Vitale, Technical Director of BioLinker.

Smart Peptides

Our team of researchers also develops smart peptides with antimicrobial effects that can be used by the food industry as an option for using the current preservatives used..

This project is carried out by Dr. Mario Pineda, Production Director at BioLinker.

To the left: image of two petri dishes containing culture medium, observing the growth of fungi.
On the right: image of the peptides in 4 glass jars.

But we will not stop here. Soon we will come up with more news!

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Importance of incubators in the growth of a Startup

Startups are companies that need incubators, have great growth potential and are newly created companies. Unlike other traditional companies, the startup enters the market before having solid capital to grow.

Image with the following description: You know what an incubator is? Know our location, emphasizing the importance of incubators.
In the background some beakers used in laboratories.

BioLinker is a startup which is located in CIETEC, which is the Innovation Center, entrepreneurship and technology.


CIETEC is a non-profit civil association, that have 20 years of existence, with the aim of “promote innovative Entrepreneurship, encouraging the transformation of knowledge into value-added products and services for the market.”

The São Paulo Technology-Based Business Incubator – USP / IPEN is the largest incubator for technology-based companies in Latin America! And its mission is, fostering innovative entrepreneurship.

CIETEC: A successful trajectory

With foundation in 1998, the incubator offers advice, mentoria e suporte na gestão tecnológica, de marketing, de busca por fomentos e de administração de micro e pequenas empresas de base tecnológica, além de uma infraestrutura física para o desenvolvimento dessa modalidade de empresa.

Por que incubar startups?

A chave dessa pergunta é inovação, e é nesse contexto que a importância das incubadoras está.

As incubadoras são organizações sem fins lucrativos que oferecem apoio físico e administrativo para as startups se estruturarem para o mercado. Can be maintained by private or public institutions, which places them as an important precursor to the culture of innovation in the university environment. Thereby, there is the generation of new ventures that could not be carried out only in Public Institutes.

Startups use technology as the basis for their innovative solution and can reach social and environmental niches that are still little explored by the market by becoming specialists in meeting a specific need..

Encouraging Startups is also important so that professionals have options beyond academic or multinational careers, increasing the possibility of professional development, mainly among scientists.

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BioTreco, applied science for children

BioTreco is a project that instigates children's curiosity about science.

This project was developed to awaken the interest in science in students from all basic education. The educational materials were developed by several USP employees and BioLinker, because we are enthusiasts of science!

Image of a girl in glasses with the title BioTreco.
At the top of the image, employees are present: BioLinker, USP, ICB USP and Institute of Biosciences.

How does it work?

In partnership with BioLinker, didactic materials were developed and kits for expression of fluorescent proteins using technology cell-free.
The project allows molecular biology assays to be carried out in the classroom, that would normally need a lot of equipment, time and money to be accomplished.
That kit allows the science teacher to demonstrate how molecular biology works in a more playful way, where students can see what was previously invisible: the proteins.

See how the expression of the fluorescent GFP protein looks:

Image of eppendorf containing fluorescent protein, with the logo.
Above, employee image:  BioLinker, USP, ICB USP and Instituto Biológico.
In the middle is written: He was curious? Contact the team!

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What is the cell-free technology used in BioLinker?

Described in 1961 the cell-free technique is still not so widespread in molecular biology laboratories, mas deveria.

Hoje vamos falar um pouco sobre esta técnica que muitos clientes acreditam que muito recente, mas na verdade, ela foi descrita na década de 60 (Nirenberg; Matthaei, 1961). O cell-free, é uma técnica que não usa a célula intacta, ela só usa a maquinaria necessária para a produção de uma proteína específica, e é por isso que nós temos um elevado rendimento.

O grande porém na época foi o elevado custo, mas a gente que é da pesquisa básica sabe: a ciência básica sempre dá um empurrãozinho na tecnologia, aí a gente tem a inovação, neh!?

E foi isso o que aconteceu…
A Mona Oliveira, PhD, no meio do seu doutorado observou que esse seria um nicho muito interessante, e ela teve a ideia de criar a BioLinker!

Desde o nascimento da BioLinker foram muitos desafios para conseguir padronizar a técnica, mas atualmente, temos o prazer de anunciar que estamos funcionando à todo vapor.

Graças a esta tecnologia, agora você pode sintetizar a sua proteína-alvo em menos de 8 horas.

Ficou curioso com as nossas atividades? Conheça os nossos produtos:

http://ip.biolinker.tech/e-commerce-proteina-sob-demanda-1841

http://ip.biolinker.tech/e-commerce-kit-covid-1111

http://ip.biolinker.tech/e-commerce-38

É uma técnica Linda e Simples:

Etapas de desenvolvimento do kit cell-free: crescimento celular, lise de células, extrato celular, adição de substrato e plasmídeo, incubação à 37 °C e obtenção da proteína alvo.
Etapas de desenvolvimento do kit cell-free. Imagem gerada em: https://biorender.com/

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