Influence of the Antibody Purification Method on Immunoassay Performance: Hapten–Antibody Binding in Accordance with the Structure of the Affinity Column Ligand

Choi, Jeongeun; Kim, Choonmi; Choi, Myung Ja

doi:10.1006/abio.1999.4261

Cited by 13 publications

(5 citation statements)

References 25 publications

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“…b Relative binding activity of the eluted antibodies (%) was measured against the original sample with the ELISA method. 58 An ELISA-elution to Dissociate Digoxin-antibody Complexes techniques [27]. It is therefore desirable to use appropriate elution conditions for the effective dissociation of the ligand from the affinity matrix with maximum yield and without affecting biological activity.…”

Section: Discussionmentioning

confidence: 99%

“…However, it has been observed that a number of parameters influence the specific elution of bound Abs when Abs are eluted against small molecules such as pesticides, drugs, etc. by using affinity purification 58 An ELISA-elution to Dissociate Digoxin-antibody Complexes techniques [27]. It is therefore desirable to use appropriate elution conditions for the effective dissociation of the ligand from the affinity matrix with maximum yield and without affecting biological activity.…”

Section: Discussionmentioning

confidence: 99%

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Application of an ELISA-elution Assay to Dissociate Digoxin-antibody Complexes in Immunoaffinity Chromatography

Lin

Wang

et al. 2010

Scandinavian Journal of Immunology

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In this study, we used a modified enzyme‐linked immunosorbent assay (ELISA)‐elution technique as a screening tool to select specific elution conditions. We examined 12 different elution conditions for the removal of antibodies from a complex on an ELISA plate; 0.2 mol/l glycine‐HCl (pH 2.5), 1.0 mol/l acetic acid (pH 2.5), 25% methanol (pH 2.5) and 3 mol/l NaSCN showed a higher elution efficiency. We conducted affinity chromatography with these four conditions for the purification of anti‐digoxin antibodies from hyperimmune sera with a digoxin‐specific column using ω‐aminoalkyl derivatives of Sepharose 4B, whose elution efficiency was similar to that of ELISA. We also monitored the relative specific activities during elution from the digoxin‐specific column. The optimum, general‐purpose dissociation reagent for this immunoaffinity system was identified as 25% methanol (pH 2.5) with an elution efficiency and relative specific activity of 88.40% and 62.25%, respectively. The high purity of the purified antibodies was demonstrated with sodium dodecyl sulphate‐polyacrylamide gel electrophoresis.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Application of an ELISA-elution Assay to Dissociate Digoxin-antibody Complexes in Immunoaffinity Chromatography

Lin

Wang

et al. 2010

Scandinavian Journal of Immunology

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“…The principle of immunoaffinity chromatography (IAC) is commonly used for purifying and concentrating nonimmunogenic small molecules (haptens) from complex samples ( − ). IAC has been used for sulfonamides ( , ), although these particular attempts have resulted in low column capacities of 2 μg sulfonamide/mL gel due to the use of nonpurified polyclonal IgG (specific and nonspecific IgG populations).…”

Section: Introductionmentioning

confidence: 99%

A Sensitive Method for Detection of Sulfamethazine and N⁴-Acetylsulfamethazine Residues in Environmental Samples Using Solid Phase Immunoextraction Coupled with MALDI-TOF MS

Sporns

2003

J. Agric. Food Chem.

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Sulfamethazine (SMT) and its major metabolite, N(4)-acetylsulfamethazine (NA-SMT), were each recovered from spiked water (0.1 ppb) and 10% (w/v) aqueous suspensions of soil (1 ppb) or composted manure (1 ppb), by using a three-stage solid phase immunoextraction (SPIE) system, followed by detection with matrix-assisted laser/desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Sulfonamide recovery rates are reported for separate stages of the SPIE system and for trace-level sulfonamide SPIE extraction from the environmental samples. SPIE MALDI-TOF MS is a rapid and definitive technique with potentially better efficiency relative to other established trace-level sulfonamide analytical methods. SPIE MALDI-TOF MS required 1.5 h per batch (8-24 samples/batch) for sample enrichment, 5 min per batch for probe preparation, and 5 min per sample to acquire and process the spectrum. This is the first time MALDI-TOF MS has been reported as a potential means of detecting trace-level drug residues in complex environmental samples.

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“…Capillary electrophoresis combined with immunoassay has been demonstrated to be a useful technique for the separation and analysis of biological compounds. − This technique allows rapid analysis, high sensitivity, small sample consumption, and easy automation. Nevertheless, proteins in the sample may be absorbed onto the capillary surface, which changes the composition and the charges of the capillary wall and detrimentally affects resolution and reproducibility. − There are usually two ways to lessen the binding of proteins to the capillary wall surface.…”

mentioning

confidence: 99%

Enhancement of the Sensitivity of a Capillary Electrophoresis Immunoassay for Estradiol with Laser-induced Fluorescence Based on a Fluorescein-labeled Secondary Antibody

Wang

Zhang

et al. 2001

Anal. Chem.

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A competitive immunoassay for estradiol (E2) based on capillary electrophoresis was established. This method was based on the competitive reaction of complete antigen and E2 with a limited amount of monoclonal antibody, with fluorescein isothiocyanate (FITC)-labeled secondary antibody as a fluorescence probe. The addition of the thermally reversible hydrogel, poly-N-isopropylacrylamide (pNIPA) in the buffer as a replaceable packing material improved reproducibility and resolution of the method. This capillary electrophoresis immunoassay with laser-induced fluorescence can be applied to determine E2 with good precision at concentrations as low as 9 pg/mL. Details of typical separations of immunological complex and free reactants are presented.

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Influence of the Antibody Purification Method on Immunoassay Performance: Hapten–Antibody Binding in Accordance with the Structure of the Affinity Column Ligand

Cited by 13 publications

References 25 publications

Application of an ELISA-elution Assay to Dissociate Digoxin-antibody Complexes in Immunoaffinity Chromatography

Application of an ELISA-elution Assay to Dissociate Digoxin-antibody Complexes in Immunoaffinity Chromatography

A Sensitive Method for Detection of Sulfamethazine and N⁴-Acetylsulfamethazine Residues in Environmental Samples Using Solid Phase Immunoextraction Coupled with MALDI-TOF MS

Enhancement of the Sensitivity of a Capillary Electrophoresis Immunoassay for Estradiol with Laser-induced Fluorescence Based on a Fluorescein-labeled Secondary Antibody

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Influence of the Antibody Purification Method on Immunoassay Performance: Hapten–Antibody Binding in Accordance with the Structure of the Affinity Column Ligand

Cited by 13 publications

References 25 publications

Application of an ELISA-elution Assay to Dissociate Digoxin-antibody Complexes in Immunoaffinity Chromatography

Application of an ELISA-elution Assay to Dissociate Digoxin-antibody Complexes in Immunoaffinity Chromatography

A Sensitive Method for Detection of Sulfamethazine and N4-Acetylsulfamethazine Residues in Environmental Samples Using Solid Phase Immunoextraction Coupled with MALDI-TOF MS

Enhancement of the Sensitivity of a Capillary Electrophoresis Immunoassay for Estradiol with Laser-induced Fluorescence Based on a Fluorescein-labeled Secondary Antibody

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A Sensitive Method for Detection of Sulfamethazine and N⁴-Acetylsulfamethazine Residues in Environmental Samples Using Solid Phase Immunoextraction Coupled with MALDI-TOF MS