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Analytical Characterization
Published in Laszlo Endrenyi, Paul Jules Declerck, Shein-Chung Chow, Biosimilar Drug Product Development, 2017
What is involved in antibody array footprinting is the synthesis of a set of short overlapping peptides (about 30) that collectively correspond to the entire amino acid sequence of a particular target biopharmaceutical. These peptides are then used to develop a set of unique polyclonal antibodies (anti-peptide antibodies) that only bind to their respective targeted peptide (which corresponds predominantly to only a small specific linear amino acid sequence region in the polypeptide chain(s) of the targeted biopharmaceutical called a linear epitope). Since many of these peptides will normally be buried within the interior of the biopharmaceutical, they will not be accessible to bind to their probing antibodies. Hence, when each of these antipeptide antibodies is separately added to a different sample of the same biopharmaceutical (using specially designed 96-well plates), it generates a unique binding profile of the antipeptide antibodies that bind to that biopharmaceutical using the sensitive and common detection capability of enzyme-linked immunosorbent assay (ELISA) technology. If another sample of the same biopharmaceutical has incurred a change in its HOS, it may trigger a change(s) in the profile of the peptides that are exposed on the biopharmaceutical’s surface in that particular sample. This HOS change would be reflected in the change in the binding profile of the antipeptide antibodies (specific to that biopharmaceutical) relative to that obtained normally from the same unaltered (native) biopharmaceutical.
Production of codon-optimized Human papillomavirus type 52 L1 virus-like particles in Pichia pastoris BG10 expression system
Published in Preparative Biochemistry & Biotechnology, 2023
Kartika Sari Dewi, Sheila Chairunnisa, Sri Swasthikawati, Dian Fitria Agustiyanti, Apon Zainal Mustopa, Wien Kusharyoto, Ratih Asmana Ningrum
B-cell epitope analysis using BepiPred Linear Epitope Prediction 2.0[20] showed that the peptide sequence located at position 476–500 (GLQARPKLKRPASSAPRTSTKKKKV) has the highest score, which is 0.709 (Figure 2a). The peptide epitope located at positions 495–500 showed the highest average score of 4.999, predicted by the Emini Surface Accessibility Prediction method (Figure 2b).[21]