Whole cell ELISA for detection of tumor antigen expression in tumor samples

Erdile, Lorne; Smith, Darlene; Berd, David

doi:10.1016/s0022-1759(01)00465-3

Cited by 28 publications

(19 citation statements)

References 8 publications

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“…A wholespore enzyme-linked immunosorbent assay (ELISA) technique was developed and optimized in our laboratory and was based on a number of whole-cell ELISA methods in which bacteria were used to coat plates at 4°C overnight (7,20,27) or at room temperature for 45 min (21). Spores were suspended in 0.03 M NaPO 4 buffer (pH 7.4) containing 4% (wt/vol) paraformaldehyde at a concentration of approximately 10 6 spores/ml.…”

Section: Methodsmentioning

confidence: 99%

Characterization of Bacillus Probiotics Available for Human Use

Duc

Hong

Barbosa

et al. 2004

Appl Environ Microbiol

385

248

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Bacillus species (Bacillus cereus, Bacillus clausii, Bacillus pumilus) carried in five commercial probiotic products consisting of bacterial spores were characterized for potential attributes (colonization, immunostimulation, and antimicrobial activity) that could account for their claimed probiotic properties. Three B. cereus strains were shown to persist in the mouse gastrointestinal tract for up to 18 days postadministration, demonstrating that these organisms have some ability to colonize. Spores of one B. cereus strain were extremely sensitive to simulated gastric conditions and simulated intestinal fluids. Spores of all strains were immunogenic when they were given orally to mice, but the B. pumilus strain was found to generate particularly high anti-spore immunoglobulin G titers. Spores of B. pumilus and of a laboratory strain of B. subtilis were found to induce the proinflammatory cytokine interleukin-6 in a cultured macrophage cell line, and in vivo, spores of B. pumilus and B. subtilis induced the proinflammatory cytokine tumor necrosis factor alpha and the Th1 cytokine gamma interferon. The B. pumilus strain and one B. cereus strain (B. cereus var. vietnami) were found to produce a bacteriocin-like activity against other Bacillus species. The results that provided evidence of colonization, immunostimulation, and antimicrobial activity support the hypothesis that the organisms have a potential probiotic effect. However, the three B. cereus strains were also found to produce the Hbl and Nhe enterotoxins, which makes them unsafe for human use.

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Section: Methodsmentioning

confidence: 99%

Characterization of Bacillus Probiotics Available for Human Use

Duc

Hong

Barbosa

et al. 2004

Appl Environ Microbiol

385

248

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“…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

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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-

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“…Thus, several thousands of samples may be handled in a short time, resulting in increased efficiency. The main fields where micotiter plates are used include high-throughput screening (HTS) (Auer, 2001;Lahana, 1999), combinatorial chemistry (Hart, 2001;Nakayama, 2001), ELISA-test (Erdile et al, 2001), and polymerase chain reaction (PCR) (Tang et al, 1999). Microbiologists are increasingly taking advantage of the mentioned properties of microtiter plates for screening of aerobic and anaerobic microorganisms.…”

Section: Introductionmentioning

confidence: 99%

Characterization of gas–liquid mass transfer phenomena in microtiter plates

Hermann

Lehmann

Büchs

2002

Biotech & Bioengineering

195

215

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Gas-liquid mass transfer properties of shaken 96-well microtiter plates were characterized using a recently described method. The maximum oxygen transfer capacity (OTR(max)), the specific mass transfer area (a), and the mass transfer coefficient (k(L)) in a single well were determined at different shaking intensities (different shaking frequencies and shaking diameters at constant filling volume) and different filling volumes by means of sulfite oxidation as a chemical model system. The shape (round and square cross-sections) and the size (up to 2 mL maximum filling volume) of a microtiter plate well were also considered as influencing parameters. To get an indication of the hydrodynamic behavior of the liquid phase in a well, images were taken during shaking and the liquid height derived as a characteristic parameter. The investigations revealed that the OTR(max) is predominantly dependent on the specific mass transfer area (a) for the considered conditions in round-shaped wells. The mass transfer coefficient (k(L)) in round-shaped wells remains at a nearly constant value of about 0.2 m/h for all shaking intensities, thus within the range reported in the literature for surface-aerated bioreactors. The OTR(max) in round-shaped wells is strongly influenced by the interfacial tension, determined by the surface tension of the medium used and the surface properties of the well material. Up to a specific shaking intensity the liquid surface in the wells remains horizontal and no liquid movement can be observed. This critical shaking intensity must be exceeded to overcome the surface tension and, thus, to increase the liquid height and enlarge the specific mass transfer area. This behavior is solely specific to microtiter plates and has not yet been observed for larger shaking bioreactors such as shaking flasks. In square-shaped microtiter plate wells the corners act as baffles and cause a significant increase of OTR(max), a, and k(L). An OTR(max) of up to 0.15 mol/L/h can be reached in square-shaped wells.

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Whole cell ELISA for detection of tumor antigen expression in tumor samples

Cited by 28 publications

References 8 publications

Characterization of Bacillus Probiotics Available for Human Use

Characterization of Bacillus Probiotics Available for Human Use

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

Characterization of gas–liquid mass transfer phenomena in microtiter plates

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