• GeNext Genomics

A Time and Cost-Effective Consistent Protein Production Platform

RESEARCH-GNG: Prashant Thakre, Chinmay Umbarkar, Komal Talreja, Sidhant Vyas, Jawad Baksh, Kalyani Dange,

Akash Nehare, Vilas Danav, Ashwin Kashikar, Supriya Kashikar and Vinod Agarkar

MARKETING-GNG: Mohammad Abdul, Rashmi Upadhyay and Supriya Kashikar


Introduction

Proteins are the most abundant macromolecules in the living system. The complexity of the molecule enhanced due

to the diversity in structural and functional properties. Protein creates dense networking interactions with nucleotide, lipid, carbohydrates and other small molecules. The living cell can contain thousands of proteins with unique role while responsible for biological functions including defense system and building blocks. However, proteins play diverse and significant role in almost all diseases such as HIV1, Tuberculosis, Influenza,

Neurodegenerative disorders, Parkinson’s disease, age related diseases and so on.

Protein functions, interactions and inhibitions are the crucial steps in the mechanisms of the living organism’s molecular processes. Decoding the characterization, network and inhibition/expression of proteins created the proteomics as the vital field or tool in life sciences. The study of broad range protein properties required for understanding the

complete set of interacting partners with the multi-functioning protein molecules.

The critical stairs in proteomic study stairways are protein extraction, purification and sample preparation. However, responsible reasons for low success rates are evaluation of posttranslational modifications and biochemical characteristics such as phosphorylation, high instability, degradation rate, unfolding and insolubility.

Generally recombinant protein expression level goes notoriously low or difficult to express and these macromolecules tend to misfold, aggregate and precipitate outside their natural environment.


Due to wide applications of recombinant proteins as therapeutic and prophylactic properties are gaining enormous requirement in the biological and biomedical industries including research activities. The high yield productions of functional and soluble proteins are the crucial goal for proteomics and protein biotechnology. Thousands of reports published on protein molecules, structures and the role in living systems. The significant fact for the success of these studies are understanding the biochemical, structural properties and in-silico predictions of the targeted protein while choosing the right heterologous protein expression vectors (modifications as per need) and systems such as E.Coli, Yeast, Insect, Mammalian etc., purification tags, media compositions, induction conditions and

purification methods. Furthermore, modification of target genes, including gene truncation, nucleotide optimization, site-specific mutations and domain specific are the key factors.


Here, TEAM-GNG successfully identified, expressed and purified the vast variety of proteins as recombinant and wild direct from the source. The range of proteins was difficult to express and purify. The domains, truncated and full proteins are available for scientific fraternities and industries as required.


Material, Method & Result

Genomic/plasmid DNA isolation, gene of interest synthesis, purification, vector preparation and cloning GNG developed own in-house protocols and methods. Modifications used wherever required if find complexity of the molecule. General time limit is 2-3 weeks including outsourced gene synthesis.




Protein expression and purification

GNG-protocols with modifications used for the expression and purification of recombinant proteins. The following Pro-1, Pro-2, Pro-5 and Pro-6 6XHIS tagged recombinant proteins supplied to industries and academics for their further studies and products. Pro-3 is the typical profile of expression and purification of recombinant proteins using metal affinity chromatography. Pro-4 protein isolated and purified directly from inclusion bodies and successfully refolded with very high yield. Pro-7 insecticidal trimetric protein expressed and purified under service contract. Pro-8 wild type >200 kDa protein isolated and purified directly from source. Pro-9 was untagged

protein purified by affinity chromatography. Pro-10 novel protease purified by ion-exchange AKTA purification for client. Pro-11 Mycobacterium and plant proteins expressed, purified and gel filtered for in-house antibody productions. General time limit is 3-4 weeks for expression, optimization and purification of recombinant proteins.




Conclusion

GeNext Genomics Pvt. Ltd. (GNG) developed as recombinant protein production platform. The services are high quality, time and cost-effective. TEAM-GNG enclosed with experienced and technically sound undergraduates, graduates and Ph.D. team mates. We produced several high value and difficult proteins for our customers and

associates. GNG is a pioneer in production of recombinant proteins as domains, truncated and full proteins. We apply following figure 7 classic model practices while dealing with the researchers needs.



Conclusion

GeNext Genomics Pvt. Ltd. (GNG) developed as recombinant protein production platform. The services are high quality, time and cost-effective. TEAM-GNG enclosed with experienced and technically sound undergraduates, graduates and Ph.D. team mates. We produced several high value and difficult proteins for our customers and

associates. GNG is a pioneer in production of recombinant proteins as domains, truncated and full proteins. We apply following figure 7 classic model practices while dealing with the researchers needs.




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