ELISA screening of monoclonal antibodies to haptens: influence of the chemical structure of hapten-protein conjugates

Danilova, N. P.

doi:10.1016/0022-1759(94)90288-7

Cited by 21 publications

(16 citation statements)

References 5 publications

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“…Digoxin was conjugated to the amino groups of microspheres by the periodate/borohydride method (44,45). It is worth noting that digoxin (MW 781) was directly linked to microspheres (44,45), while a haptenic derivative (2) including a spacer group had to be used in the assay for theophylline (M r 180).…”

Section: Resultsmentioning

confidence: 99%

“…It is worth noting that digoxin (MW 781) was directly linked to microspheres (44,45), while a haptenic derivative (2) including a spacer group had to be used in the assay for theophylline (M r 180). The digoxin molecule is large enough to occupy most of the antibody binding site; thus, a spacer arm between the digoxin structure and the carrier surface was not necessary to improve the digoxin assay (45).…”

Section: Resultsmentioning

confidence: 99%

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A Duplexed Microsphere-Based Fluorescent Immunoassay

Szurdoki

Michael

Walt

2001

Analytical Biochemistry

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

A Duplexed Microsphere-Based Fluorescent Immunoassay

Szurdoki

Michael

Walt

2001

Analytical Biochemistry

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“…Free digoxin concentration was measured by competitive solid-phase immunoenzyme assay [2] 5 min and 30 rain postinjection. To this end, 100 gl serum and 100 gl standard digoxin solutions with concentrations of 0.01, 0.05, 0.1, 0.2, 0.5, and 1 ~tg/ ml were dialyzed for 4 h against 300 gl physiological saline so that the data obtained with serum samples containing only digoxin and digoxin and ADA were comparable.…”

Section: Methodsmentioning

confidence: 99%

Effect of antidigoxin monoclonal antibodies on cardiac disturbances caused by high concentrations of digoxin

et al. 1998

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Effect of high-affinity mouse monoclonal antidigoxin antibodies D22 on digoxin-induced cardiac disturbances is studied in guinea pigs injected with 0.5 mg/kg digoxin. Five minutes after digoxin injection, bradycardia, isolated extrasystoles, conduction disturbances, and changes in the ST segment and QRS complex appear; digoxin concentration attains 0.2-0.4 gg/ml. Antidigoxin monoclonal antibodies injected 10 min after digoxin slightly reduce heart rate, prevent isolated extrasystoles, improve atrioventricular conduction, and normalize the amplitude of the ST segment and the shape of the QRS complex. Serum digoxin concentration decreases to 0.015-0.03 gg/ml. These data suggest that monoclonal antidigoxin antibodies can be effectively used in acute digoxin intoxication.

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“…The inherent problems associated with producing MAbs to small haptens are well documented and discussed (Clementi et al, 1991;Chappey et al, 1994;Danilova, 1994;Mittenbuhler et al, 1997;Tuomola et al, 2000). In an attempt to successfully produce hapten-specific MAbs, we have examined both in vivo and in vitro immunisation protocols using different methods of hapten conjugation to carrier molecules and various detection/screening methods.…”

Section: Discussionmentioning

confidence: 99%

Methods for generation of monoclonal antibodies to the very small drug hapten, 5-benzimidazolecarboxylic acid

Morán

O’Keeffe

O’Connor

et al. 2002

Journal of Immunological Methods

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Drug-specific monoclonal antibodies (MAbs) were produced against the very small drug hapten (162.15 Da), 5-benzimidazolecarboxylic acid, an analogue of 2-(4-Thiazolyl)benzimidazole (TBZ) but lacking the thiol group. TBZ is widely used as a broad-spectrum anthelmintic in various animal species and humans and also as a food preservative and agricultural fungicide. The anti-5-benzimidazolecarboxylic acid antibodies produced have potential use for extraction and/or detection of protein-bound residue forms of TBZ.Three in vivo immunisation regimes (with combinations of two related small drug haptens and two different adjuvants/ carrier molecules) and an in vitro immunisation procedure using a combination of three related unconjugated small drug haptens were investigated. Specificity for the hapten immunogen/s was initially determined using two different ELISA procedures. BIACORE analysis, in conjunction with drug binding inhibition studies, was used to confirm the specificity of a small number of selected clones.In vivo immunisation with a drug molecule conjugated to a lipopeptide/T-cell epitope, which acts both as a carrier molecule and an adjuvant was the most useful of the methods tested for the production of specific MAbs to a typically very small hapten with low immunogenic properties.

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ELISA screening of monoclonal antibodies to haptens: influence of the chemical structure of hapten-protein conjugates

Cited by 21 publications

References 5 publications

A Duplexed Microsphere-Based Fluorescent Immunoassay

A Duplexed Microsphere-Based Fluorescent Immunoassay

Effect of antidigoxin monoclonal antibodies on cardiac disturbances caused by high concentrations of digoxin

Methods for generation of monoclonal antibodies to the very small drug hapten, 5-benzimidazolecarboxylic acid

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