| Popis: |
Reversed-phase, high pressure liquid chromatography (HPLC) in a modified TFA/H2O-TFA/acetonitrile gradient has been successfully applied to the structural analysis of serologlcally defined rabbit immunoglobulln heavy chains. This mobile phase modification yields a relatively flat baseline at high UV sensitivity. Approximately 40 distinct tryptic pep-tides were resolved from each heavy chain, representing the VHa+ (a), a2, and a3) and VHa− (y33,30 and y33,−) immunoglobulin allotypes. About 30 peptides were shown to be derived from the Fc region (CH2 and CH3), and 8–10 peptides from the Fd region (VH1 and CH1). Seven Fd peptides were shared by all VHa+ and VHa− heavy chains. The al and a2 digests each displayed one allotype-specific peptide, whereas no allotype-specific peptides were observed for the a3 heavy chain. No differences were detected between the y33,30 and y33,− peptides; however, both expressed a common y-specific pep-tide. Amino acid analysis of purified al-specific and y-specific pept... |
