Solutions

Large molecules are being used to assist in treating conditions such as cancers, immune diseases, and other infectious diseases. Our platform covers the entire early-stage research in the large molecule discovery and carries it further to drug development.

At Stellixir, we offer a wide range of integrated discovery biology services to meet your drug discovery needs. Our services include Recombinant DNA technology, protein sciences, cell-based assays, DMPK, toxicology, and other in vitro assays.

Recombinant DNA Technology:

We support and offer a diverse 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.

Cloning and Vector Generation:

• 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:

Plasmid DNA purification service caters to both small research facilities as well as large-scale manufacturing for biotech and 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.

Mutagenesis:

• Customized mutagenesis requirements such as random and site-directed mutagenesis and mutant library generation.
• Point mutation, insertions, deletion or gene fragment replacements.
• Gene shuffling, combinatorial mutagenesis, alanine scanning and site saturation mutagenesis.

Antibody Gene Sequencing and Reformatting:

Antibody gene sequencing and reformatting at Stellixir is a one-stop solution for recombinant expression and production of antibodies.

• RACE (Rapid Amplification of cDNA Ends) using template switching adapter ligation.
• Degenerate 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:

• 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: Constructing cDNA libraries, phage libraries and yeast libraries along with provisions for high throughput screening and functional characterization.

Gene Expression Analysis:

Scientists at Stellixir carry out routine expression studies by several methods;

• 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 that remain in a recombinant biotherapeutics. We ensures that these impurities and contaminants are detected and reduced to levels below those guided by regulatory agencies.

• Quantitative Real time PCR
• ELISA
• SPR (Biacore)
• Mycoplasma test

Protein Sciences

We offer a comprehensive range of activities from sequence optimization to final product generation (Gene to Protein). We offer the following services and solutions with the flexibility of multiple entry points to help our clients.

Our Protein Sciences Services Include:

Protein Expression:

We offer

• Designing, scouting and screening studies for challenging proteins.
• Protein expression studies.
• Scale-up using spinner flask, shaker flask, HYPER flask and WAVE bioreactor platforms to handle volumes up to 50 L scale with low operating costs.

Mammalian Expression Systems:

• Mammalian systems used for expression are CHO, ExpiCHO, HEK, ExpiHEK, etc.
• Protein production in transient expression mode or using stable clones or pools.

Insect Cell Expression Systems:

Baculovirus mediated protein expression and production in Sf9, Sf21, Hi5 and super Sf9 insect cell lines

Yeast Expression Systems:

Stable protein expression in Pichia, recombinant molecule is typically secreted in media by genome integration.

Bacterial Expression Systems:

• Soluble, periplasmic and inclusion bodies in E. coli
• Multi-protein expression by co-transfection or duet plasmid technology
• Refolding condition screening and method optimization

Protein Purification:

We are proficient in purifying tagged as well as un-tagged and all classes of target proteins from mammalian, yeast and microbial expression systems.

• Purification process optimization
• Protein purification in batch binding mode as well as automated FPLC systems
• Chromatographic purification methods include IMAC, IEC, mixed-mode, HIC, SEC and affinity for micrograms to gram-scale quantities

Antibody Purification:

• Purification of monoclonal antibodies of different origins (Human, non-human Primates, Mouse, Rat, Rabbit, Pig, Hamster, Gerbil, etc.)
• Purification of antibody fragments of various formats (Fab, Fab2, F(ab)’2, VHH, scfv and generation from full length mAbs
• Bi-specific and KIH antibodies
• Diabodies
• Single domain antibody
• Protein purification from hybridoma cultures

Microbial Purification Capabilities:

• Soluble protein
• Periplasmic protein
• Intracellular protein
• Refolding from inclusion bodies
• Purification of Major Histocompatibility (MHC) complexes
• Membrane protein production - GPCRs, nanodisc technology, etc.
• Antibody drug conjugate screening, development and purification
• High throughput protein screening and purification
• Protein cleavage and purification

Protein Analytics and Characterization

We are committed to deliver high-quality products by performing a number of routine protein analyses throughout all project stages ensuring the identity, purity, structural and conformational integrity, stability and function of the protein.

• Quantitative analysis: Protein estimation using Absorbance at 280nm, Bradford and BCA assays
• Qualitative analysis: SDS-PAGE (Reducing/Non-reducing) and DOT/Western blot analysis
• Purity: SE-HPLC, IEX-HPLC and RP-HPLC
• Protein stability analysis (Real-time and Accelerated Stability Testing)
• LC-MS analysis
• Capillary electrophoresis (CE).
• Peptide Mass Fingerprinting (PMF)
• Biological activity and binding studies using enzymatic assays, ELISA, Biacore, etc.
• Endotoxin analysis
• Host cell protein (HCP) and Host cell DNA (HCD) analysis

Protein Stability Analysis

• Real-time and accelerated stability testing.
• Long-term stability studies.
• Purity: SE-HPLC, IEX-HPLC and RP-HPLC

Protein Conjugation and Labelling

• Protein labelling/conjugation (HRP, biotin, Ruthenium, fluorescent labels, etc.)
• Conjugation of resin with specific protein/peptide