Development and optimization of a cell-based neutralizing antibody assay using a sample pre-treatment step to eliminate serum interference

McCutcheon, Krista; Quarmby, Valerie; Song, An

doi:10.1016/j.jim.2010.03.016

Cited by 15 publications

(3 citation statements)

References 9 publications

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“…Chromatography on immobilized Protein A/G has recently been used successfully as a pretreatment procedure to eliminate non-specific assay interferences in individual serum samples from rheumatoid arthritis patients 62 (see Appendix for sample protocol).…”

Section: Immunoadsorption Chromatography On Immobilized Igg Binding Pmentioning

confidence: 99%

“…Protein A/G is a gene fusion product that contains four Fc binding domains from Protein A and two from Protein G. 61 As it binds to all human IgG subclasses, it can be particularly helpful in purifying IgG antibodies where the subclass identity is not known. Chromatography on immobilized Protein A/G has recently been used successfully as a pretreatment procedure to eliminate non-specific assay interferences in individual serum samples from rheumatoid arthritis patients 62 (see Appendix for sample protocol).…”

Section: Immunoadsorption Chromatography On Immobilized Igg Binding Proteinsmentioning

confidence: 99%

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Analytical error and interference in immunoassay: minimizing risk

2011

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Although generally robust, immunoassays remain vulnerable to occasional analytical errors that may have serious implications for patient care. Sporadic errors that occur as a result of properties of the specimen are particularly difficult to detect. They may be due to the presence of cross-reacting substances, antianalyte antibodies or antireagent antibodies, all of which may lead to erroneously high or low results. Low results may be observed for tumour markers due to high-dose hooking in the presence of very high analyte concentrations. Erroneous results can occur unexpectedly with any specimen and there is no practical means of identifying specimens likely to cause problems in immunoassays. The possibility of interference should always be considered when results do not appear to be in accord with the clinical picture. Errors can occur in even the best-managed laboratories and their early investigation is always desirable. If there is any doubt whatsoever about a result, clinical staff should be encouraged to contact the laboratory. Investigations for possible interference that can be undertaken in most laboratories include testing for linearity on dilution, recovery experiments, treatment with heterophilic blocking tubes and confirmation using a different method. It may be desirable to consult specialist laboratories if more complex studies are necessary. Informing clinical and laboratory staff of the everpresent possibility of unexpected interference, ensuring brief clinical details are available to laboratory staff, and above all facilitating excellent communication between laboratory and clinical staff are key to minimizing the risk of clinical mismanagement due to unsuspected interference.

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Section: Immunoadsorption Chromatography On Immobilized Igg Binding Pmentioning

confidence: 99%

Section: Immunoadsorption Chromatography On Immobilized Igg Binding Proteinsmentioning

confidence: 99%

Analytical error and interference in immunoassay: minimizing risk

2011

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“…Although very sensitive, cell-based assays are more prone to interfering processes from sample matrix. To circumvent this problem, strategies to eliminate the disturbing factors by sample pre-treatment have been developed 22 23 . These strategies enable the detection of nAbs even in the presence of high antigen levels in serum.…”

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confidence: 99%

Detection of Infertility-related Neutralizing Antibodies with a Cell-free Microfluidic Method

Eyer

Root

Verboket

et al. 2015

Sci Rep

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The unwanted emergence of neutralizing antibodies (nAbs) against an endogenous or a therapeutic protein can result in deficiency diseases or therapy failure. Here, we developed a cell-free microfluidic method for the sensitive detection and quantification of nAbs in human serum that are associated with infertility. We used cell-derived vesicles containing the luteinizing hormone (LH)/choriogonadotropin receptor (LHHCGR) to detect nAbs against LH. The method exploits the entire cellular signal amplification mechanism, and facilitates the detection of as little as 0.44 nM of LH-nAb (Kd 1.5 nM) in human serum matrix within only 15 minutes. In addition, dose-response curves can be generated in less than 2 hours to evaluate the nAB concentration and dissociation constant. The developed system is devoid of problems associated with cell-based assays and we believe that this simple effect-directed analysis can be used in clinical environments, and is adaptable to other hormones or cytokines and their respective nAbs.

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Standardization and Validation of Cell‐Based Assays for the Detection of Neutralizing Anti‐Drug Antibodies

Finco‐Kent

Grenham

2011

Detection and Quantification of Antibodies to Biopharmaceuticals

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Development and optimization of a cell-based neutralizing antibody assay using a sample pre-treatment step to eliminate serum interference

Cited by 15 publications

References 9 publications

Analytical error and interference in immunoassay: minimizing risk

Analytical error and interference in immunoassay: minimizing risk

Detection of Infertility-related Neutralizing Antibodies with a Cell-free Microfluidic Method

Standardization and Validation of Cell‐Based Assays for the Detection of Neutralizing Anti‐Drug Antibodies

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