Enzyme-immunoassays for polychlorinated biphenyls: structural aspects of hapten-antibody binding

被引:26
作者
Franek, M [1 ]
Pouzar, V [1 ]
Kolar, V [1 ]
机构
[1] ACAD SCI CZECH REPUBL, INST ORGAN CHEM & BIOCHEM, CR-16610 PRAGUE 6, CZECH REPUBLIC
关键词
immunoassays; immunogens; tracers; PCBs; molecular models; interactions; assay characterization;
D O I
10.1016/S0003-2670(97)00318-8
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
The syntheses of 4,4'-dichlorobiphenyl- and 3',4,4',5-tetrachlorobiphenyl-2-azo-protein immunogens and the corresponding haptens conjugated to peroxidase through the amidoadipate spacer arm are described. Only one sheep of the 24 immunized animals yielded an antiserum with superior antibody titres. The direct competitive enzyme-linked immunosorbent assay (ELISA) incorporating this antiserum showed IC50 values for commercial polychlorinated biphenyl (PCB) mixtures Delor 103, 104, 105, and 106 (the corresponding formulations are Aroclor 1242, 1248, 1254 and 1260) of about 200, 250, 600, and 900 mu g l(-1), respectively. No cross-reactions with chlorinated phenols, phenoxyalkanoic acids, or chlorinated herbicides were observed and only a 5% cross-reactivity was found for DDE (Delor 103=100%). Molecular models of hapten determinants were constructed to examine potential conformational changes in the biphenyl nucleus in relation to tracer immunoreactivity and assay selectivity. The structural data were not consistent with the lack of immunoreactivity for peroxidase tracers in conjunction with almost all of the tested antisera. Influence of the various molecular factors on the binding and competitive characteristics is discussed.
引用
收藏
页码:163 / 176
页数:14
相关论文
共 27 条
[1]   Binding of phenylphosphocholine-carrier conjugates to the combining site of antibodies maintains a conformation of the hapten [J].
Barbar, E ;
Martin, TM ;
Brown, M ;
Rittenberg, MB ;
Peyton, DH .
BIOCHEMISTRY, 1996, 35 (09) :2958-2967
[2]   SYNTHESIS OF A FLUORESCENT ANALOG OF POLYCHLORINATED-BIPHENYLS FOR USE IN A CONTINUOUS-FLOW IMMUNOSENSOR ASSAY [J].
CHARLES, PT ;
CONRAD, DW ;
JACOBS, MS ;
BART, JC ;
KUSTERBECK, AW .
BIOCONJUGATE CHEMISTRY, 1995, 6 (06) :691-694
[3]   A MONOCLONAL IMMUNOASSAY FOR THE COPLANAR POLYCHLORINATED-BIPHENYLS [J].
CHIU, YW ;
CARLSON, RE ;
MARCUS, KL ;
KARU, AE .
ANALYTICAL CHEMISTRY, 1995, 67 (21) :3829-3839
[4]  
DURHAM LJ, 1963, ORG SYNTH, V4, P555
[5]   CHARACTERIZATION OF ANTIBODIES AGAINST ESTRONE-AZO-PROTEIN CONJUGATES USING H-3 AND I-125 RADIOLIGANDS [J].
FRANEK, M ;
HRUSKA, K .
JOURNAL OF STEROID BIOCHEMISTRY AND MOLECULAR BIOLOGY, 1983, 19 (03) :1363-1370
[7]   DEVELOPMENT OF A MICROCOLUMN RADIOIMMUNOASSAY FOR SCREENING OF POLYCHLORINATED-BIPHENYLS IN MILK AND IN ANIMAL FATS [J].
FRANEK, M ;
HRUSKA, K ;
SISAK, M ;
DIBLIKOVA, I .
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 1992, 40 (09) :1559-1565
[8]   GLUTARAMYL-BETA-ALANYL SPACER GROUP FOR HAPTENIC COUPLING TO PROTEINS - PREPARATION OF IMMUNOGENS FOR ANTIBODY-PRODUCTION AGAINST POLYCHLORINATED-BIPHENYLS [J].
GOON, DJW ;
NAGASAWA, HT ;
KEYLER, DE ;
ROSS, CA ;
PENTEL, PR .
BIOCONJUGATE CHEMISTRY, 1994, 5 (05) :418-422
[9]   RAPID ANALYSIS OF PCBS IN SOIL BY ENZYME-IMMUNOASSAY [J].
HARRISON, RO ;
MELNYCHUK, N .
INTERNATIONAL JOURNAL OF ENVIRONMENTAL ANALYTICAL CHEMISTRY, 1995, 59 (2-4) :179-185
[10]  
JEFFREY PD, 1995, J MOL BIOL, V248, P344, DOI 10.1006/jmbi.1995.0226