Comparison of pH and ionic strength dependence of interactions between monoclonal antibodies and bovine beta-lactoglobulin.
A panel of 13 monoclonal antibodies (mAbs) against distinct determinants on bovine beta-lactoglobulin, a model protein antigen, were examined and compared for their ability to bind and desorb from the antigen at differing pHs and ionic strengths by an enzyme-linked immunosorbent assay and elution assay. Among them, mAb 61C1 was found to be highly sensitive to the pH, and 3 in 4 mAbs directed to the region 42-56 also strongly depended on the change in ionic strength. Because of the large proportion of charged amino acid residues in the region 42-56, the electrostatic forces are considered to be more predominant than the hydrophobic interactions in the latter antigen-antibody reactions, thereby resulting in their high sensitivity to the ionic strength.
Kamata
N
N
Department of Biological and Chemical Engineering, Gunma University, Japan.
Enomoto
A
A
Ishida
S
S
Nakamura
K
K
Kurisaki
J
J
Kaminogawa
S
S
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000937:Antigen-Antibody Reactions; D001665:Binding Sites; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D006433:Hemocyanins / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007782:Lactoglobulins / Q000378:metabolism*; D008969:Molecular Sequence Data; D009994:Osmolar Concentration
D000911:Antibodies, Monoclonal; D007782:Lactoglobulins; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin