Job opening in Affigenix – Chennai

Affigenix Biosolutions Private Limited is hiring.

Qualification: Undergraduate and Master’s degree from Biochemistry, Microbiology and Biotechnology field.

Responsibility: Development of novel diagnostic kits.

Location: Diagnostic kit manufacturing facility located at Alathur near Chennai, Tamil Nadu.

Check Affigenix website for more information: www.affigenix.com.

If interested, please share your updated CV with cover letter to info@affigenix.com.

Bio Patrika interviews Dr. Sengupta on her thoughts about “directionality of proteolysis”

Dr. Bhaswati Sengupta’s interview with Bio Patrika hosting “Vigyan Patrika“, a series of author interviews. Dr. Sengupta is currently working on Nucleosome dynamics and chromatin remodelling using smFRET as a postdoc in the Department of Chemistry at Pennsylvania State University, USA. She did her PhD from the lab of Prof. Pratik Sen, Department of Chemistry, Indian Institute of Technology Kanpur, India. She published last part of her PhD work titled “Fluorescence correlation spectroscopy as a tool to investigate the directionality of proteolysis” as the first author in International Journal of Biological Macromolecules (2020).

How would you explain your paper’s key results to the non-scientific community?

Proteins are a long chain of amino acid molecules connected by a peptide bond. Proteolysis is a significant part of the digestion system where the protein gets cleaved into smaller peptides. In this paper, we have used human serum albumin (HSA) as a model protein and papain, chymotrypsin, and trypsin as enzymes to digest HSA. Being a long-chain protein, HSA can be cleaved from either side. In this work, we have used fluorescence correlation spectroscopy (FCS) to understand exactly from which side these enzymes cleave HSA. FCS enables us to measure the change in the size of the fluorescently labelled fragments. As the digestion proceeds, the size becomes smaller, which we measure by FCS. By preferentially labelling to two sides of the protein chain with fluorescent dyes, we can see from which side the digestion process starts. The principle of the measurement is shown in Scheme 1.

Scheme 1: Monitoring protein digestion by single molecule technique.

HSA is a large protein having three domains. For HSA, we found out that digestion starts from the domain I for all three enzymes under consideration. Fluorophore 1 (TMR) slows the faster decrease of size as compared to fluorophore 2 (NPCE) labelled one, which indicates that the fragment size decreases faster for the domain I indicating that digestion starts from the domain I. We have also performed reverse-phase high-performance liquid chromatography (RP-HPLC) to cross-check the findings, supporting our results.

What are the possible consequences of these findings for your research area?

Several techniques enable the measurement of digested fragments after proteolysis like mass spectrometry, HPLC, etc. The uniqueness of the proposed approach is its ability to tell directionality using a minimum amount of protein. FCS is sensitive and can be successfully used for picomolar to nanomolar sample concentrations. This is a new application of the single-molecule research that can be used to understand proteins’ digestion profile.

“[…] I see you are now able to think of novel ideas and design the experiments; your training has been completed”

What was the exciting moment (eureka moment) during your research?

There is more than one Eureka moment associated with this specific work. This was the last project I worked on during my PhD. When I conceived the idea and shared with my boss, he said, “I see you are now able to think of novel ideas and design the experiments; your training has been completed”. That was some words of motivation before I could start the next venture. When I started working, I was unsure if I would see any difference in the digestion profile monitored by two different fluorophores labelled at two other locations. If there was no difference, there was no specific conclusion, and it wouldn’t have been as fascinating as the current finding. So when I observed a difference in the digestion profile, I was pretty excited, which was definitely the second eureka moment. The third eureka moment would be when the reverse phase HPLC supported the single-molecule measurements. That was the moment when we found proof that whatever we see in single-molecule measurement is valid.

What do you hope to do next?

Regarding the piece of work, I talked about, I am not following up on that anymore. We had several ideas of trying the technique on different proteins and used different enzymes and different environments. Also, to use drugs to see if they affect the digestion profile etc. But unfortunately, I had to leave to join a postdoc and couldn’t continue with it. If someday I get a chance, I would like to work on the extended ideas. 

Where do you seek scientific inspiration?

The best (or the worst) thing being a researcher is that I don’t know what waits ahead, and from my early childhood, I have felt tremendous attraction towards what is unknown. It feels exciting to delve into the strange world, which is why I keep loving science. Besides, I am blessed to have supervisors (PhD and postdoc) who motivate and have always believed in my ability and dedication. The excitement of discovering something new is often inexplicable. All these factors keep me going, and I wish to be roaming around the field of science forever.

How do you intend to help Indian science improve?

Honestly, “improving Indian science” is a huge goal, and I don’t think I have reached the stage to think about it. Right now, I am doing postdoctoral research and wish to set up my lab in India someday. Provided I can do that; my next goal would be to design experiments with minimum budget and resources possible given the poor condition of the academic budget in India. The next target would be to equip the lab with efficient and budget-friendly instruments. Once all these are in place, one can use single-molecule experiments for several biological and chemical applications using a minimum sample amount. This will cut down the cost of buying or producing expensive samples again and again.

Reference

Sengupta B, Das N, Singh V, Thakur AK, Sen P. Fluorescence correlation spectroscopy as a tool to investigate the directionality of proteolysis. Int. J. Biol. Macromol. 2020; 164: 2524-2534.

Email: bhaswati1487@gmail.com

Job opening in Biocon Biologics

BIOCON Biologics is Hiring.

Department: Biosimilars – MSAT (Downstream)

Experience: 7-12 years

Qualification: MSc/MTech/PhD

Responsibilities:

  1. To manage MSAT Lab functions on routine basis for Downstream related activities.
  2. To ensure product life cycle management.
  3. To handle continuous verification of process, deviation and out of specifications.
  4. To evaluate the alternate vendors for critical raw materials and consumables as required.
  5. Contribute at team discussions of CMC and other decision-making forums for development, clinical and commercial manufacturing.
  6. Review Technology transfer of manufacturing process to CMOs (outside parent site).
  7. Scientific knowledge on microbial downstream processes.

Interested candidates with relevant experience may share profile at sushmitha.101@biocon.com.

Job opening in Kemwell Biopharma

Kemwell Biopharma is hiring experienced professional for Bangalore location.

Position: Executive, Quality control (QC)

Skills required: Experience in in-vitro Bioassays and potency determining ELISA

Experience: 1-3 years

Requirement: Immediate

If you are interested, please share your updated resume to debatree.chakrabarty@kemwellpharma.com

Job openings in CuraTeQ Biologics

Six job openings in CuraTeQ Biologics – Hyderabad.

Skill required: Cell culture, purification and analytical sciences functions.

Qualification: PhD in relevant study area, pursued post-doctoral research for a period of at least 2 years and/or 2-3 years of post- PhD industry work. Overseas PhDs from top 30 Universities (QS Ranking) can apply without post- doctoral experience.

If you are interested, please share your updated resume to HR Nobby Paul (nobby.paul@curateqbio.com).

Job openings in Biocon Biologics – Chennai

Multiple job openings in R&D department, Biocon Biologics – Chennai.

Skill required: Cell Culture, MAb’s, well versed with R&D upstream process, Mammalian cell culture techniques, Relevant experience in R& D process.

Working experience in Biologics/Biosimilar/Biopharma industry.

Years of experience: 8 to 12 years

Qualification: Ph.D/Master’s in Biotechnology

Location: Chennai

If you are interested, please share your updated resume to shilpa.BBHRDCON11@biocon.com

Job opening in Aurigene

Department: Cells & Molecular Biology

Work Location: Bangalore

No. of position: 2

Qualification: M.Sc. with 3-5years’ experience

Key responsibilities:  

  • To work as part of drug discovery project teams in both internals and collaboration programs.
  • Conduct cell based assays and molecular biology experiments using primary cells and cell lines.
  • Establish various high throughput in-vitro assays for screening of molecules such as proliferation assay, ELISA, Western Blot analysis, In-cell-western assay, reporter assay, qPCR etc.
  • Experience with target validation experiments using techniques such as knock down using siRNA, shRNA & CRISPR is an added advantage.
  • Able to troubleshoot experiments, analyse data and prepare scientific presentations.
  • Independently design and execute experiments and present data to cross functional teams.

Interested candidates with relevant experience may send their resume to preeti_5@aurigene.com.

For more details, please call at +91-9590737048.

Job opening in docplexus

Job opening for Senior medical writer in docplexus

Qualification: MBBS, BHMS, BAMS, PhD and Post-graduate degree in lifesciences discipline

Experience: 3+ years

Requirements:

  • Excellent writing, editing and organizational skills.
  • An excellent understanding of the pharmaceuticals industry and drugs. development process.
  • Ability to manage multiple writing projects at one time.
  • Ability to work independently as well as in a team environment.

Interested candidates with relevant experience may send their resume to neha.shingote@docplexus.in.

Bio Patrika interviews Dr. Baidya on his thoughts about “role of key phosphorylation sites in GPCRs”

Dr. Mithu Baidya’s interview with Bio Patrika hosting “Vigyan Patrika“, a series of author interviews. Dr. Baidya is currently a postdoc in the lab of Prof. Arun Shukla in the department of Biological Sciences and Bioengineering at the Indian Institute of Technology Kanpur, India. He published a paper titled “Key phosphorylation sites in GPCRs orchestrate the contribution of β‐Arrestin 1 in ERK1/2 activation” as the first author in EMBO reports (2020).

How would you explain your paper’s key results to the non-scientific community?

Our research published in EMBO reports explains the molecular basis of side effects caused by drugs, targeting a special type of receptor called G protein coupled receptors (GPCRs). Almost 40% of current drugs target these GPCRs for treating several clinical conditions. Therefore, the study has enormous implications in designing the next generation of safer drugs having very minimal side effects.

GPCRs are membrane proteins that act as a mailbox that receives several messages in the form of hormones or drugs and relays the information from the external environment to the cell machinery for a response in a coded massaging system inside the cell, which we call as signaling (Figure 1). GPCRs have two main partners who serve as a pivotal point for disbursing the signal viz. G proteins and beta-arrestins. GPCRs, upon receiving signal/massage undergo a modification called phosphorylation i.e., the addition of phosphate group on Serine and Threonine amino acids present in their terminal tail. This phosphorylation serves to act as a code that beta-arrestins recognizes and binds the receptor and initiates signaling in the cells.

Figure 1: GPCR behaves like a mailbox that receives messages from outside the cell and relays them to cell machinery by signaling.

 Typically, GPCR signaling involves ERK1/2 MAP kinase proteins, which play an essential role in cellular growth and proliferation. For most of the GPCRs, lowering the level of beta-arrestins typically reduces ERK1/2 activation, but for some GPCRs, it leads to an enhancement. This phenomenon of opposing contribution of beta-arrestin in different receptors was inexplicable with our current understanding. It was quite a puzzling mystery in the field for more than two decades.

In this study, we took two such GPCRs vasopressin 2 receptor and bradykinin 2 receptor which is known to have an opposing effect in ERK activation upon beta-arrestin depletion. To unclutter this puzzle, we designed a smart and simple experimental approach whereby we changed the arrangement of the code i.e. position of Serine and Threonine amino acids in the tail of the human bradykinin receptor, a GPCR that is involved in cardiovascular regulation and pain sensation. By even a slight rearrangement of the code in bradykinin receptor C tail, we observed a dramatic alteration in the inhibitory contribution of beta-arrestin 1 in ERK1/2 activation to a more supportive role. We further discovered that such a change in the receptor C tail code does not affect the overall physical binding of beta-arrestin 1 to the bradykinin receptor but it rather changes its conformation (i.e. overall shape of beta-arrestin 1), which in turn determines beta-arrestin’s ability to support ERK1/2 activation in the cells (Figure 2).

Figure 2: Conformation of beta-arrestin (βarr) in vasopressin 2 receptor stimulates ERK signaling but a different beta-arrestin conformation in bradykinin 2 receptor is inhibitory to ERK activation but when Bradykinin 2 receptor mutant with a changed code, made similar to vasopressin 2 receptor now induce a conformation in beta-arrestin which again becomes supportive of ERK activation.

What are the possible consequences of these findings for your research area?

More than one third of all marketed medicines target the GPCRs to treat a wide variety of ailments. Understanding the details of GPCR signaling and regulation has critical implications for making smarter medicines with a minimized side-effect. Therefore, our findings now provide a previously missing piece of information about how conformation (overall shape) of beta-arrestins can control GPCR signaling, and therefore, have the potential to contribute towards improving GPCR targeting medicines in the future.

“[…] a paradigm-changing contribution to the field to understand beta-arrestin plasticity and related functions better.”

What was the exciting moment (eureka moment) during your research?

Conformation drives specific functions of beta-arrestin 1, which is a big finding of the study. Our preliminary study showed that beta-arrestin 1 exists in distinct conformation in different GPCRs first established in vasopressin and bradykinin receptor systems. It turned out to be a ‘Rosetta stone’ to decrypt this opposing beta-arrestin function in the other receptors. This is a paradigm-changing contribution to the field to understand beta-arrestin plasticity and related functions better. I feel this was the inflection point in the whole study, which made us so upbeat to follow this up to its logical conclusion.

What do you hope to do next?

Our next big aim is to identify the conformations of beta-arrestin, which favors biased signaling focused more towards the GPCR-beta-arrestin axis in different GPCRs, which will help us generate next-generation drugs with much lesser side effects in many more GPCRs. Our focus will be more towards those GPCR whose alternation is known to cause critical diseases in humans.

Where do you seek scientific inspiration?

Getting a new piece of data that crystallizes my hypothesis (in many cases opposing it) is a treat of delight, and this unsettles me (in the right sense, of course) and gives me the drive to chase it for months to come. Finally, when peers appreciate these discoveries makes me feel I am moving in the right direction.

How do you intend to help Indian science improve?

I am not sure how I can improve Indian science, but I want to enhance or intend to improve the pharmaco-profile in the context of side effects caused by current GPCR drugs. By understanding the finer details of the beta-arrestin signaling paradigm, we can design much safer drugs with lesser side effects, which will bring relief to patients undergoing treatments with GPCR drugs. GPCR research has not picked up in India the way it should have been, especially keeping in mind India’s massive production output in global pharmaceutical manufacturing. This is quite a paradox really, and can be explained by the fact India does not discover or identify new GPCR drugs but rather produce the generic ones for foreign labels. This has to improve and can only be done by a higher focus in GPCR research in academia and the pharma industry. Let’s hope we only improve in the coming years.

Reference

Baidya M, Kumari P, Dwivedi-Agnihotri H, Pandey S, Chaturvedi M, Stepniewski TM, Kawakami K, Cao Y, Laporte SA, Selent J, Inoue A, and Shukla AK. Key phosphorylation sites in GPCRs orchestrate the contribution of β-Arrestin 1 in ERK1/2 activation. EMBO Rep. 2020; 21(9): e49886.