Reduction of matrix interferences by the combination of chaotropic salt and DMSO in a broadly applicable target-based ELISA for pharmacokinetic studies of therapeutic monoclonal antibodies

Doucet, Julie; Canadi, Jasna; Kalis, Christoph; Valentin, Marie‐Anne; Marrony, Séverine; Deckert-Salva, Fabienne; Legay, François; Avraméas, Alexandre

doi:10.1016/j.jpba.2009.06.029

Cited by 6 publications

(3 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The endogenous TEX101 protein present in pooled seminal plasma was used to calibrate the assay. Because limit of detection (LOD) of the immunoassay without seminal plasma pretreatment was not high enough, we examined literature (15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25) to identify conditions that would improve the assay sensitivity in seminal plasma. Thus, seminal plasma was subjected to pretreatment with the following detergents, salts and solvents: 1% SDS, 2-5% Triton X-100, 1% CHAPS, 2% sodium deoxycholate, 1 M urea, 3 M guanidine hydrochloride, 10% dimethyl sulfoxide, 10% dimethylformamide, 10% glycerol, 10% methanol, 10% acetonitrile.…”

Section: Production Purification and Analysis Of Recombinant Human mentioning

confidence: 99%

Immunocapture-Selected Reaction Monitoring Screening Facilitates the Development of ELISA for the Measurement of Native TEX101 in Biological Fluids*

Korbakis

Brinc

Schiza

et al. 2015

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

Monoclonal antibodies that bind the native conformation of proteins are indispensable reagents for the development of immunoassays, production of therapeutic antibodies and delineating protein interaction networks by affinity purification-mass spectrometry. Antibodies generated against short peptides, protein fragments, or even full length recombinant proteins may not bind the native protein form in biological fluids, thus limiting their utility. Here, we report the application of immunocapture coupled with selected reaction monitoring measurements (immunocapture-SRM), in the rapid screening of hybridoma culture supernatants for monoclonal antibodies that bind the native protein conformation. We produced mouse monoclonal antibodies, which detect in human serum or seminal plasma the native form of the human testis-expressed sequence 101 (TEX101) protein-a recently proposed biomarker of male infertility. Pairing of two monoclonal antibodies against unique TEX101 epitopes led to the development of an ELISA for the measurement of TEX101 in seminal plasma (limit of detection: 20 pg/ml) and serum (limit of detection: 40 pg/ml). Measurements of matched seminal plasma samples, obtained from men pre-and post-vasectomy, confirmed the absolute diagnostic specificity and sensitivity of TEX101 for noninvasive identification of physical obstructions in the male reproductive tract. Measurement of male and female serum samples revealed undetectable levels of TEX101 in the systemic circulation of healthy individuals. Immunocapture-SRM screening may facilitate development of monoclonal antibodies and immunoassays against na-

show abstract

Section: Production Purification and Analysis Of Recombinant Human mentioning

confidence: 99%

Immunocapture-Selected Reaction Monitoring Screening Facilitates the Development of ELISA for the Measurement of Native TEX101 in Biological Fluids*

Korbakis

Brinc

Schiza

et al. 2015

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

show abstract

“…Matrix interference by serum proteins is a common finding during capture ELISA development and may be ascribed to a variety of reasons. Potential causes include the cross-linking of the capturing and detecting mAbs or the binding of the target Ag to serum components, thus preventing its interaction with one or both mAbs (13,25).…”

Section: Serum Compatibility Of the Mycolactone Capture Assaymentioning

confidence: 99%

“…However, diluting the sample may drive the mycolactone concentration below the limit of detection of the assay, and performing lipid extraction for every sample is an added complication. A more straightforward way of dealing with matrix interference is to use suitable chaotropic agents that are capable of breaking the (typically) low-affinity interactions of the interferents (25). We modified our assay buffer by adding different concentrations of MgCl 2 , MgSO 4 , or ammonium thiocyanate, all of which are commonly used chaotropic agents.…”

Section: Serum Compatibility Of the Mycolactone Capture Assaymentioning

confidence: 99%

An Antigen Capture Assay for the Detection of Mycolactone, the Polyketide Toxin of Mycobacterium ulcerans

Warryn

Dangy

Gersbach³

et al. 2021

The Journal of Immunology

View full text Add to dashboard Cite

Mycolactone is a cytotoxin responsible for most of the chronic necrotizing pathology of Mycobacterium ulcerans disease (Buruli ulcer). The polyketide toxin consists of a 12-membered lactone ring with a lower O-linked polyunsaturated acyl side chain and an upper C-linked side chain. Mycolactone is unique to M. ulcerans and an immunological Ag capture assay would represent an important tool for the study of Buruli ulcer pathogenesis and for laboratory diagnosis. When testing sets of mycolactone-specific mouse mAbs, we found that Abs against the hydrophobic lower side chain only bind mycolactone immobilized on a solid support but not when present in solution. This observation supports previous findings that mycolactone forms micellar structures in aqueous solution with the hydrophobic region sequestered into the inner core of the aggregates. Although an Ag capture assay typically requires two Abs that recognize nonoverlapping epitopes, our search for matching pairs of mAbs showed that the same mAb could be used both as capture and as detecting reagent for the detection of the mycolactone aggregates. However, the combination of a core-specific and a core/upper side chain–specific mAb constituted the most sensitive ELISA with a sensitivity in the low nanogram range. The results of a pilot experiment showed that the sensitivity of the assay is sufficient to detect mycolactone in swab samples from Buruli ulcer lesions. Although the described capture ELISA can serve as a tool for research on the biology of mycolactone, the assay system will have to be adapted for use as a diagnostic tool.

show abstract

Use of response surface methods and path of steepest ascent to optimize ligand-binding assay sensitivity

Joyce¹,

Leung²

2013

Journal of Immunological Methods

View full text Add to dashboard Cite

Reduction of matrix interferences by the combination of chaotropic salt and DMSO in a broadly applicable target-based ELISA for pharmacokinetic studies of therapeutic monoclonal antibodies

Cited by 6 publications

References 21 publications

Immunocapture-Selected Reaction Monitoring Screening Facilitates the Development of ELISA for the Measurement of Native TEX101 in Biological Fluids*

Immunocapture-Selected Reaction Monitoring Screening Facilitates the Development of ELISA for the Measurement of Native TEX101 in Biological Fluids*

An Antigen Capture Assay for the Detection of Mycolactone, the Polyketide Toxin of Mycobacterium ulcerans

Use of response surface methods and path of steepest ascent to optimize ligand-binding assay sensitivity

Contact Info

Product

Resources

About