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Biology |
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Inbiopro provides one expert source for 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.
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Custom Antibody & Stable Cell Line Development |
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Inbiopro provides everything necessary for the creation and characterization of stable cell lines expressing a gene of interest at high and homogeneous levels. The cell culture scientists focus on optimizing all parameters specific to the requisite cell line to efficiently produce stable cell lines according to requirements. The scientists at Inbiopro have extensive experience in developing stable cell lines using CHO, HEK, NSO for biotherapeutic manufacturing.
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Transient Transfection |
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Inbiopro’s 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.
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Assay Development & Screening |
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Inbiopro’s assay development process is based on stringent quality criteria and comprehensive controls. The scientific team has extensive experience in developing new assays depending on the client’s needs. Inbiopro's screening capabilities span from biochemical to cell-based functional assays.
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Analytical Services |
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Here proteins based on size are separated and are 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.
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Protein Characterization |
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Western: This technique permits identification of proteins by their molecular weight and their immunoreactivity
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IEF: 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.
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Glycan Analysis: 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.
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Circular dichroism (CD): Circular dichroism (CD), is the differential absorption of left- and right-handed circularly polarized light which arises dye 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 characterising 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 the effect of other additives or interactions such as ligand binding on protein structure.
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N-Terminal Sequencing: N-Terminal Sequencing is a tool for the sequencing / determination of the amino acid residues (from the N-terminus) of a protein or peptide.
N-terminal Sequencing 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
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MALDI-TOF: MALDI-TOF (Matrix Assisted Laser Desorption Time of Flight) 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
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Host Cell Protein: Recombinant biopharmaceuticals can be contaminated by residual host cell proteins (HCP). 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.
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Host Cell DNA: 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.
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Endotoxin Test: 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.
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SDS Page: Here proteins based on size are separated and are 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.
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HPLC |
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RP HPCL: 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.
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SEC HPCL: 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.
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Trytic Peptide Analysis: 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.
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