Dual microcolumn immunoaffinity liquid chromatography: an analytical application to human plasma proteins.
Dual-column microcolumn immunoaffinity chromatography was accomplished by coupling immunoaffinity (IAC) and reversed-phase high-performance liquid chromatography (RP-HPLC) analysis columns. The IAC support was prepared by hydrophobically adsorbing the appropriate antibody onto an octyl silica-based stationary phase contained in a 250-microns-l.d. fused-silica microcolumn. Once the antigen was captured, the antigen-subtracted human plasma could be sent to the RP-HPLC microcolumn for plasma profiling. The antigen was quantitated by desorbing it from the IAC column and transferring it to the RP-HPLC system. Human plasma profiles were generated after removal of albumin, and transferrin and alpha 1-antitrypsin were quantitated with nanogram sensitivity. All analyses were accomplished with less than 1 microL of human plasma.
Flurer
C L
CL
Department of Chemistry, Indiana University, Bloomington 47405.
Novotny
M
M
D000418:Albumins / Q000737:chemistry; D000936:Antigen-Antibody Complex; D001798:Blood Proteins / Q000032:analysis* / Q000302:isolation & purification; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D014168:Transferrin / Q000032:analysis; D000515:alpha 1-Antitrypsin / Q000032:analysis
D000418:Albumins; D000936:Antigen-Antibody Complex; D001798:Blood Proteins; D014168:Transferrin; D000515:alpha 1-Antitrypsin