Our state-of-art facility houses a molecular biology department with highly qualified scientists to support and offer a wide range of services in the areas of recombinant DNA technology, antibody gene sequencing, mutagenesis, library generation, plasmid DNA preparation, microbial genome engineering, gene expression analysis, and IVT mRNA synthesis.
Recombinant DNA Technology
- Cloning and vector generation
We support you with our gene cloning service to customize the generation of recombinant constructs such as antibodies, enzymes, and reagent proteins. Our methods include:- Ligation dependent and independent cloning
- Recombination based high throughput cloning
- De novo cloning (Synthetic gene fragments, PCR and cDNA based)
- Customized vector construction based on client requirement
- Plasmid DNA preparation
Our plasmid DNA purification service caters to both small research facilities, biotechs and larger pharmaceutical companies.- Research grade plasmid at small scale suitable for cloning, mutagenesis, microbial transformation and protein production
- High quality industrial grade plasmid suitable for protein, antibody production, stable cell line generation in mammalian cells, bacmid generation for protein expression in insect cells, viral packaging, in vitro mRNA synthesis, vaccine, gene therapy studies and animal immunization
- Process development for customized plasmid propagation and purification
- Large scale manufacturing
- Mutagenesis
We offer site-directed mutagenesis services and support our clients by creating specific DNA mutations to study gene regulation, DNA-protein interactions, protein structure/function relationship, enzyme function, and novel recombinant proteins.- Point mutation, insertions, deletion, or gene fragment replacements
- Customized mutagenesis requirements such as random and site-directed mutagenesis and mutant library generation
- Gene shuffling, combinatorial mutagenesis, alanine scanning, and site saturation mutagenesis.
Antibody Gene Sequencing and Reformatting
Antibody sequence information from hybridoma is essential for protecting, preserving, and optimizing monoclonal antibodies (mAbs). Antibody gene sequencing and reformatting at Aurigene is a one-stop solution for recombinant expression and production of antibodies. Our offering includes:
- RACE (Rapid amplification of cDNA ends) using template switching adapter ligation
- Degenerated oligos
- De novo antibody sequencing and analysis using LC-MS/MS
- Reformatting antibody gene sequences into various other production formats like bispecifics, mAb, scFv, Fab, F (ab)'2, VHH/nanobody
- Antibody gene sequencing for mouse, rat, rabbit and other species
Strain engineering
Our experienced team can support you wtih genome-scale modification of bacterial and yeast strains for the production of recombinant proteins, platform chemicals, and metabolic engineering.
- BSL-1 and BSL-2 microbes
- Gene knock-out and knock-in using homologous recombination-based methods
- Characterization of engineered strains using microbiological methods, RT-PCR, and advanced analytics
- CRISPR-Cas9 based mutagenesis
Library generation and evaluation
Our clients have access to our cDNA, phage, and yeast libraries which we apply for high throughput screening and functional characterizations.
Gene expression analysis
Regulation and expression of genes at transcriptional and translational stages play a crucial role in determining gene functions. We can carry out routine expression studies by several methods including:
- Quantitative real-time PCR
- ELISA in various formats
- Flow cytometry
- Western blotting
- Mass spectrometry
Residual protein, DNA and mycoplasma detection
Residual host-cell proteins (HCP) and host-cell DNA (HCD) are impurities in recombinant biotherapeutics, which need to be detected and quantified to prove that they meet the limits set by global regulatory agencies. To do that, we apply the following methods:
- Quantitative real-time PCR
- ELISA
- SPR (Biacore)
- Mycoplasma test
Aurigene Pharmaceutical Services is
one of the leading contract research organization (CRO)
provides the best contract research manufacturing
FAQ
There are two ways to get the sequence of an antibody. One is by polymerase chain reaction (PCR) and the other is by Mass Spectrometry (MS). The first method used either RACE (Rapid Amplification of c-DNA Ends) method, where RNA extracted from an antibody-producing cell is converted to c-DNA, amplifying the antibody genes by PCR, and cloning the gene Sanger sequencing and match with available databases to decipher the antibody sequence. The second method employs digestion of the antibody using proteolytic enzymes followed by measuring the mass of the digested peptide fragments by MS and comparing it across databases to find the amino acid sequence of the antibody.
Yes, it can.
Mutagenesis can be increased by using several chemical additives, proper strains of bacteria and media, and varying physical parameters, thereby increasing the stringency of the process to enrich the library.
Ribonuclease H (RNaseH) is a family of non-sequence-specific endonuclease enzymes that catalyze the cleavage of RNA in an RNA/DNA substrate via a hydrolytic mechanism. Since c-DNA involves the formation of RNA/DNA complex, RNaseH will degrade the parent RNA strand resulting in termination/failure of the DNA amplification process from RNA, thereby affecting gene cloning.
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