An immunoassay for the detection and quantitative determination of mycothiol.
Mycothiol (MSH) is a glycosylated derivative of N-acetylcysteine that may have antioxidant functions in mycobacteria and other actinomycetes. To develop a highly specific assay for MSH, we capitalized on the selective binding of thiols to a maleimide residue linked to bovine serum albumin and employed affinity-purified polyclonal antibody and an enzyme-linked secondary antibody for detection. The assay was shown to be specific and to detect MSH at levels as low as 0.1 pmol when conducted in the form of a microtiter plate-based ELISA. A similar, nitrocellulose membrane-based immunoassay was shown to be useful for qualitative detection of MSH-producing bacterial colonies.
Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla 92093, USA.
M D
MD
Unson
G L
GL
Newton
C
C
Davis
R C
RC
Fahey
D000818:Animals; D000906:Antibodies / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D003545:Cysteine; D004187:Disaccharides / Q000032:analysis*; D013293:Enterococcus faecalis / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000378:metabolism; D005260:Female; D006020:Glycopeptides; D006433:Hemocyanins / Q000737:chemistry; D007074:Immunoglobulin G / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification; D007150:Immunohistochemistry; D007294:Inositol; D009161:Mycobacterium / Q000378:metabolism; D011720:Pyrazoles; D011817:Rabbits; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000737:chemistry; D013295:Streptococcus mutans / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000032:analysis*
D000906:Antibodies; D004187:Disaccharides; D006020:Glycopeptides; D007074:Immunoglobulin G; D011720:Pyrazoles; D013438:Sulfhydryl Compounds; C089265:mycothiol; D012710:Serum Albumin, Bovine; D007294:Inositol; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin; D003545:Cysteine