A competitive radioimmunoassay for peptidoglycan monomer.
The preparation and characterization of the essential components to be used in the radioimmunoassay of peptidoglycan monomer (PGM) is described. In order to raise the anti-peptidoglycan monomer antibodies 14C-labelled peptidoglycan monomer-bovine serum albumin conjugate was prepared by the coupling of 14C-peptidoglycan monomer to bovine serum albumin in the presence of glutaraldehyde in 0.1 M NaHCO3 at pH 8.3. The prepared conjugate elicited anti-PGM response in rabbits. A synthetic analog of peptidoglycan monomer, Boc-L-tyrosyl-peptidoglycan monomer was prepared by condensation of unprotected peptidoglycan monomer and N-hydroxysuccinimidester of Boc-L-tyrosine in the presence of triethylamine and the obtained disaccharide-hexapeptide was labelled with Na125I. This compound exhibited the ability of binding to anti-peptidoglycan monomer antibodies. The prepared compounds, namely anti-PGM antibodies and 125I-labelled Boc-L-tyrosyl-peptidoglycan monomer, were used as essential components in competitive radioimmunoassay for peptidoglycan monomer determination in mammalian and human sera and plasma, respectively.
Vranesić
B
B
Department of Radioimmunology, Institute of Immunology, Zagreb, Croatia, Yugoslavia.
Ljevaković
D
D
Tomasić
J
J
Ladesić
B
B
D000595:Amino Acid Sequence; D000906:Antibodies; D001951:Brevibacterium; D002240:Carbohydrate Sequence; D002250:Carbon Radioisotopes; D002850:Chromatography, Gel / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D002855:Chromatography, Thin Layer / Q000379:methods; D004590:Electrophoresis, Paper / Q000379:methods; D008969:Molecular Sequence Data; D010457:Peptidoglycan / Q000032:analysis* / Q000302:isolation & purification; D011863:Radioimmunoassay / Q000379:methods
D000906:Antibodies; D002250:Carbon Radioisotopes; D010457:Peptidoglycan