Stem Cell-based Therapeutics

Yashraj Biotechnology Ltd. stem cell-based therapeutics program is aimed at leveraging our proprietary induced pluripotent stem cell (iPSC) and mesenchymal stem cell technology to troubleshoot and counter challenging medical conditions. With the ability to play a special role in the repair and maintenance of the human body, stem cell-based therapeutics are playing a critical part in revolutionizing the therapeutics field. The prospect of stem cell-based regenerative medicine as an alternative to conventional drug-based therapies is becoming a tangible reality by the day owing to the vigorous commitment of the research communities in studying the potential applications across a wide range of diseases. Stem cells offer the perfect solution when there is a need for tissue and organ transplantation through their ability to differentiate into the specific cell types that are required for repair of diseased tissues. At Yashraj Biotechnology, we make use of our extensive dataset of stem cell technology acquired from preclinical studies along with our team of well qualified researchers to advance human health and regenerative medicine.

Non-Stem Cell-based Therapeutics

Cell-based therapy as a modality of regenerative medicine is considered one of the most promising disciplines in the fields of modern science & medicine. Recent research reporting successful translation of cell therapies to patients have enriched the hope that such regenerative strategies may one day become a treatment for a wide range of vexing diseases. In fact, the past few years witnessed a rather exponential advancement in clinical trials revolving around cell-based therapies. The therapeutic potential of somatic cell therapy has been more extensively investigated, particularly the anti-cancer potential of immune cell transfer. Living cells also constitute a crucial component in the development of therapeutic products, not only in the bio-pharmaceutical industry development but also in traditional drug discovery programs for e.g. cell-based assays are used in small molecule drug discovery to gather information on biological activity, toxicology, molecular mechanisms and off-target effects. Cell-based assays are also critical for immune system monitoring, including effector cell activation, immune checkpoint modulation, and cytokine signaling.

At Yashraj Biotechnology Ltd, we offer patient-derived primary and patient iPSC-derived differentiated cells that can be used for cell-based therapy regime for a patient. We have built expertise in many of the capabilities required to successfully realize the promise of cell-based therapies including patient sample sourcing, gene editing and differentiation technology. We apply expertise in standardization, quality assurance and knowledge of GMP manufacturing, freeze/thaw technology and formulation to move from the lab to manufacturing cells for clinical use. We continue to evolve our capabilities to create and deliver value-added products & services. We collaborate with partners who have expertise and capabilities complementary to our own in order to accelerate the development of our cell-based therapies.

Gene Therapy

Gene therapy is a tool used for modifying a person’s genes as a therapeutic modality in patients for curing the disease. Gene therapies can work by introducing the vectors that carry healing gene and the mechanism could be by (a) replacing a disease-causing gene with a healthy one; (b) inactivating a disease-causing gene that is constitutively active (eg. Oncogene); and (c) introducing altogether a new or modified gene into the body to help treat a disease.

YBL has established facilities and a validated & robust procedure for production of components used in Gene Therapy such as Plasmid DNA (non-viral vectors) and Adeno Associated Virus (AAV) and Lentivirus (Viral vectors). We also have validated procedures to edit, such as introducing a gene or inactivating or deleting a gene, in Patient derived autologous cells. YBL would like to work closely and collaborate with Industry and Academia in providing End-to-End services in developing gene therapy products and perform preclinical testing for Safety, Efficacy, toxicity, and to determine the maximum tolerance dose using mice. By this, YBL is committed to its mission of making a healthier world and a passionate dedication to the patients most in need of these therapies.

Viral vectors

Viral vectors are a modified version of a virus that is being targeted to deliver therapeutic gene(s) into our cells. There are several types of viral vectors including lentivirus and adeno-associated virus (AAV) each with its own advantages and disadvantages for specific applications. Although many viral vectors are safe and effective delivery vehicles for clinical gene therapy, some viral vectors are considered risky for potential oncogenes.

AAV vectors have been frequently used for gene therapy applications due to the lack of pathogenicity and toxicity, ability to infect both dividing and non-dividing cells, and long-term transgene expression profiles. Lentiviral vectors, specifically the 3rd and 4th generation have become a popular tool for gene transfer experiments and gene therapy approaches, specifically when stable gene transfer is required, resulting in durable transgene expression by integrating into the host cell genome.

At YBL, a highly robust AAVs manufacturing process is established which yields 1014 to 1015 TU/mL of GMP grade AAVs. All necessary QC tests will be performed with a CoA. The whole process is designed and regulated under QA supervision. YBL also has the robust methods established for large scale production of Lentivirus at 108 to 109 TU/mL which are used for CAR-T cell production and other R&D purposes.

Non-viral vectors

In Gene therapy, Non-viral vectors are used to transfer nucleic acids inside the cell as an alternative for enzyme/ protein/ antibodies, includes Plasmid DNA vectors and messenger RNA (mRNA). The optimal vector and delivery system depends on the target cells, duration of expression and the size of the genetic material to be incorporated in the vector.

In Gene Therapy, the Plasmid DNA represents the simplest form of vector for transport of DNA across cell nuclei for expression of therapeutic protein. Plasmids are easy to handle and stable at ambient temperature with a very large DNA packaging capacity.

mRNA based gene therapy represents a promising alternative because of their high potency, capacity for rapid development and potential for low-cost manufacture and safe administration; including easy manipulation, rapid and transient expression, and adaptive convertibility without mutagenesis, which are suitable for therapeutic modalities.

YBL has the state of the art facility including Bioreactors to synthesize GMP grade plasmid DNA (CCC- covalently closed circular topology) and mRNA (nascent and 5’ capped) in small to medium to large quantities for R&D as well as for commercial purpose. We provide End-to-End solutions from design to supply.

Gene Knock-in

Gene knock-in involves the targeted insertion of an exogenous gene/ gene-of-interest at a specific locus in the genome to modify the function of an endogenous gene for studying human disease modeling and effects of genes with altered functions. Gene Knock-in is safer and more reliable compared to random integration due to targeted insertion of gene(s) by Homology Directed Repair (HDR) in the known locus.

At YBL, CRISPR-Cas9 technology is well established for Targeted Gene Knock-in in human cell lines (both autologous and allogeneic). We use Non-viral (plasmid DNA) based genes for targeted insertion of gene(s) and developing knock-in cell lines. We provide End-to-End services from designing to development of single cell colonies to expansion and supply of cell lines.

Gene Knock-out

Gene knockout involves the total removal or permanent deactivation of a gene that is achieved by genetic engineering. Several molecular biology techniques were developed for getting Gene Knock-out including siRNA, shRNA, ZNF, TALENs and lately the CRISPR/Cas system. The CRISPR/Cas system works by editing genes by precisely recognizing and cutting DNA and cellular DNA repair processes will induce gene deletion. The system consists of two key parts namely the Cas9 enzyme, an endonuclease and a guide RNA, a sequence that recognizes the target site

YBL has well established robust and thoroughly validated processes for developing knockout cell lines using CRISPR/Cas technology with spCas9, eCas9 and nCas9. YBL provides End-to-End services for developing gene knockout in cell lines of choice from designing of sgRNA sequences, cloning, single cell colony isolation of knockout cells and validation.