Studies on the heterogeneity of anti-hapten antibodies by means of two-dimensional affinity electrophoresis

✍ Scribed by Professor Kazusuke Takeo; Tatehiko Tanaka; Kazuyuki Nakamura; Ryosuke Suzuno

• Publisher: John Wiley and Sons
• Year: 1989
• Tongue: English
• Weight: 758 KB
• Volume: 10
• Category: Article
• ISSN: 0173-0835
• DOI: 10.1002/elps.1150101204

✦ Synopsis

The molecular heterogeneity of rabbit anti-hapten antibodies has been investigated by two-dimensional affinity electrophoresis (2D-AEP). Anti-dansyl and antiarsanilic diazo-antibodies were separated into several hundred IgG spots as in the case of anti-DNP antibodies. They were grouped into a number of monoclonal IgG families. At the beginning of immunization, IgG spots having low pland low affinity were predominant but one or two weeks after immunization the IgG spots with high affinity and high p l increased. After the second or third immunization, the 2D-AEP patterns became stable and constant. Anti-arsanilic diazo and anti-DNP antibodies exhibited only weak cross-reactivity with other aromatic haptens. In contrast, antidansyl antibodies cross-reacted to a considerable degree with DNP-hapten. A few anti-dansyl IgG families which have cross-reactivity with DNP-hapten were separated. Their apparent dissociation constants to the haptens and their affinity ratios were calculated from their 2D-AEP patterns, according to the affinity theory.