Kappa marker typing with high-performance liquid chromatography: identification of kappa marker specific tryptic peptide from the kappa light chain of immunoglobulin G.
Human serum immunoglobulin G was separated into its heavy and light chains by sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred electrophoretically to a polyvinylidenedifluoride membrane. Peptide fragments liberated from the light chain by in situ digestion with trypsin were then analyzed by reversed-phase high-performance liquid chromatography (HPLC). On comparing the HPLC patterns of these fragments derived from three major kappa marker (Km) types, two distinct peaks specific for the Km types were detected. Sequencing of the two specific peak peptides confirmed that they were identical to a stretch comprising residues 191-207 of the immunoglobulin kappa light chain, which contains valine/leucine allotypic variation at position 191.
Iida
R
R
Department of Legal Medicine, Fukui Medical School, Japan.
Yasuda
T
T
Nadano
D
D
Takeshita
H
H
Kishi
K
K
D000595:Amino Acid Sequence; D015415:Biomarkers; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis* / Q000302:isolation & purification; D007145:Immunoglobulin kappa-Chains / Q000032:analysis* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D014357:Trypsin
D015415:Biomarkers; D007074:Immunoglobulin G; D007145:Immunoglobulin kappa-Chains; D014357:Trypsin