Discovery and characterization of a peptide motif that specifically recognizes a non-native conformation of human IgG induced by acidic pH conditions.
In therapeutic antibody preparation, acidic pH conditions are generally used for elution from Protein A affinity column of IgG or for its viral inactivation. Exposing IgG to low pH conditions induces conformational changes, leading to its functional damage or loss, although the mechanisms have not been fully elucidated. In this study using random peptide T7 phage display libraries, we isolated a unique and novel peptide motif that specifically recognized the non-native conformer (acid conformer) of human IgG that was generated by the low pH treatment, but not the native conformer. We examined the generation conditions and biochemical properties of acid conformer using the peptide motif as an affinity ligand. The acid conformer was easily generated at acidic pH (<pH 3.0) and at moderate temperatures (20-40 degrees C). The conformer was present in a monomeric form functionally maintaining antigen or Fc receptor binding, but showed a tendency to aggregate with a long incubation time at neutral pH (>25 degrees C). The peptides isolated here could contribute to the elucidation of the mechanisms of antibody dysfunction or aggregation during acid exposure as well as storage of human IgG.
Sakamoto
Kotaro
K
Faculty of Engineering, Kagoshima University, Korimoto, Kagoshima 890-0065, Japan.
Ito
Yuji
Y
Hatanaka
Takaaki
T
Soni
Preeti Brijiral
PB
Mori
Toshiyuki
T
Sugimura
Kazuhisa
K
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000737:chemistry*; D008024:Ligands; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry*; D011485:Protein Binding; D011487:Protein Conformation; D011961:Receptors, Fc / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D013696:Temperature
D007074:Immunoglobulin G; D008024:Ligands; D019151:Peptide Library; D010455:Peptides; D011961:Receptors, Fc