Dual microcolumn immunoaffinity liquid chromatography: an analytical application to human plasma proteins |
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Authors: | CL Flurer M Novotny |
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Affiliation: | Department of Chemistry, Indiana University, Bloomington 47405. |
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Abstract: | 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. |
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