Inbiopro provides a comprehensive range of solutions and competencies to clients based on their specific requirements.
Inbiopro offers reliable and economical molecular biology outsourcing, including de novo gene synthesis, gene cloning, DNA construct, plasmid preparation, and site directed mutagenesis. The services cover the entire range of gene analysis from identification and cloning to functional studies.
Inbiopro provides niche, cutting-edge tools for the creation and characterization of stable cell lines expressing a gene of interest at high and homogeneous levels. Our cell culture scientists have extensive experience in developing stable cell lines using CHO, HEK, and NSO for biotherapeutic manufacturing. They focus on optimizing all parameters specific to the requisite cell line to efficiently produce stable cell lines.
Our transient mammalian expression system has been used to produce a diverse range of products. Typical results achieve high concentrations of harvestable and active products. Large-scale transient transfection is carried out using HEK 293 cells in serum-free conditions.
Inbiopro’s assay development process is based on stringent quality criteria and comprehensive controls. The scientific team has extensive experience in developing new assays focused on the client’s needs. Our screening capabilities span from biochemical to cell-based functional assays.
- Here proteins based on size are separated and used to determine sample purity. Proteins in the polyacrylamide gel are detected using either Coomassie blue or silver stain for more sensitive detection. Inbiopro's assay provides a measure of sample purity as well as an approximate molecular weight.
- This technique permits identification of proteins by their molecular weight and their immunoreactivity.
- Isoelectric focusing separates proteins based on their charge and can detect heterogeneity in samples due to the presence of post-translational modifications such as glycosylation. The IEF gel can be visualized using either Coomassie blue or silver stain for more sensitive detection. Inbiopro's assay provides an approximation of the pI of a protein sample as well as an assessment of the extent of heterogeneity in the sample.
- The glycosylation sites of a therapeutic protein product are analyzed and reported by peptide mapping service. All the common forms of N-glycosylation in therapeutic antibodies produced by CHO cells (including G0F, G1F, G2F, G3F, G4F, and G5F) can be routinely identified by the peptide mapping service. In this, the N-glycan is first released by peptide N-glycosidase (PNGase), and then derivatized by a reaction with 2-AB. A normal phase HPLC with a fluorescence detector is used to profile all N-glycan forms.
- Circular Dichroism (CD) is the differential absorption of left- and right-handed circularly polarized light, which arises due to the asymmetry of naturally occurring molecules such as proteins and sugars. The CD spectrum is the wavelength dependency of the difference in absorption between the right and left-handed components. CD spectroscopy is particularly good for characterizing protein secondary and tertiary structure, for comparing the structures of proteins from different manufacturing processes, formulations or mutations, and for studying the effects of temperature, pH, denaturants and other additives or interactions such as ligand binding on protein structure.
- N-terminal sequencing is a tool for the sequencing/ determination of the amino acid residues (from the N-terminus) of a protein or peptide. It may be employed for:
- Identity determination of the product or impurity;
- Evaluation of potential truncation of the N-terminus;
- Evaluation of product purity/ degradation (as evidenced by multiple N-termini);
- Proteomics support for protein / peptide identification.
- Matrix Assisted Laser Desorption Time of Flight (MALDI-TOF) Mass Spectrometry is one of the most specialized mass spectrometric techniques available. MALDI-TOF-MS analysis gives fast, accurate data that can provide the following information:
- Peptide/ protein molecular weight;
- Sequence of peptides;
- Analysis of a peptide during a protease digestion experiment at selected time intervals.
- Recombinant biopharmaceuticals can be contaminated by residual HCPs. These proteins produced by the host cell during cell culturing or fermentation can cause an immune response in patients at levels as low as 10 to 100 parts per million (ppm). Prior to the approval of a biological product for therapeutic use, the level of contaminating host cell proteins in the product must be quantitatively measured according to the ICH and FDA guidelines.
- Products isolated from biological sources are potentially contaminated by nucleic acids derived from the host cell system or other components used during cultivation and purification.
- Inbiopro offers endotoxin analysis of raw materials, finished products, and pharmaceutical water and steam samples, using the Limulus Amoebocyte Lysate (LAL) test by the kinetic-turbidimetric instrumental method. The kinetic-turbidimetric method is an instrumental technique, which monitors the reaction rate of the sample with LAL reagent.
- Reversed-phase HPLC offers a purity analysis that is not dependent upon size or charge as are other common methods for determining purity. RP-HPLC can efficiently separate similar proteins based on hydrophobicity and will provide a quantitative measure of protein impurity in a sample.
- Size exclusion HPLC analysis separates proteins based on their molecular size. An assay can detect protein aggregation in a sample down to 1% using SEC HPLC with UV detection.
- In the tryptic peptide analysis, samples are reduced and alkylated, then subsequently digested with trypsin. The resulting tryptic peptide mixture is analyzed by reversed-phase HPLC with UV detection. The tryptic peptide mapping analysis provides a qualitative profile or map of the peptides generated from a trypsin digest of a sample.