Immunochromatographic membrane strip assay system for a single-class plasma lipoprotein cholesterol, exemplified by high-density lipoprotein cholesterol measurement.
In assessing risk factors of coronary heart disease, a membrane immunochromatographic system that minimizes requirements of instrument and reagent handling was investigated by utilizing high-density lipoprotein (HDL) cholesterol (HDL-C) as model analyte. The system is composed of four functional membrane strip pads connected in sequence as follows (from the bottom): immunoseparation based on the biotin-streptavidin reaction; catalytic conversion of cholesterol to hydrogen peroxide; production of a colorimetric signal; and induction of a continuous wicking of medium. For immunochromatography, a monoclonal antibody, specific to apolipoprotein B100 that is present on the surfaces of low-density lipoproteins (LDL) and very low-density lipoproteins (VLDL), with a high binding constant (5 x 10(10) L/mol), was raised and chemically conjugated to streptavidin. The conjugate was first reacted with lipoprotein particles, and this mixture was absorbed by the capillary action into the biotin pad of the system. After being transferred by medium, immunocapture of LDL and VLDL particles onto the biotin pad took place, and in situ generation of a colorimetric signal in proportion to HDL-C occurred consecutively. The capture was selective as well as effective (minimum 88% of LDL and VLDL in clinical concentration ranges), and the detection limit of the HDL-C was far lower than 20 mg per 100 mL. The same concept may also be applicable to LDL cholesterol measurement provided suitable antibodies specific to HDL and VLDL are available.
Paek
S H
SH
Graduate School of Biotechnology, Korea University, Seoul 136-701, Korea.shpaek@tiger.korea.ac.kr
Jang
M R
MR
Mok
R S
RS
Kim
S C
SC
Kim
H B
HB
D000818:Animals; D000911:Antibodies, Monoclonal; D001709:Biotechnology; D001710:Biotin; D008076:Cholesterol, HDL / Q000032:analysis* / Q000097:blood / Q000276:immunology; D002845:Chromatography / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods; D008074:Lipoproteins / Q000032:analysis / Q000097:blood; D051379:Mice; D019809:Streptavidin
D000911:Antibodies, Monoclonal; D008076:Cholesterol, HDL; D008074:Lipoproteins; D001710:Biotin; D019809:Streptavidin