A cell surface ELISA for the screening of monoclonal antibodies to antigens on viable cells in suspension

Grunow, Roland; D’Apuzzo, Massimo; Wyss‐Coray, Tony; Frutig, Karin; Pichler, Werner J.

doi:10.1016/0022-1759(94)90232-1

Cited by 24 publications

(11 citation statements)

References 15 publications

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“…The receptor binding event is essentially based on the direct hFasRECD recognition function of hFasLECD in native states. Thus, the applications are considered to be suitable for the fluorescent detection of cell-surface hFasR in viable cells, which can eliminate the background and the false positive reactions derived from the modifications of cell surface caused by fixation [32, 33]. Concerning the modifications with functional proteins, the possible conjugation with Fab’ fragments and other related domains of monoclonal antibodies specific to surface antigens will provide an opportunity for the targeting of hFasLECD to diseased cells.…”

Section: Discussionmentioning

confidence: 99%

Site-specific chemical conjugation of human Fas ligand extracellular domain using trans-cyclooctene – methyltetrazine reactions

Muraki

Hirota

2017

BMC Biotechnol

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BackgroundFas ligand plays a key role in the human immune system as a major cell death inducing protein. The extracellular domain of human Fas ligand (hFasLECD) triggers apoptosis of malignant cells, and therefore is expected to have substantial potentials in medical biotechnology. However, the current application of this protein to clinical medicine is hampered by a shortage of the benefits relative to the drawbacks including the side-effects in systemic administration. Effective procedures for the engineering of the protein by attaching useful additional functions are required to overcome the problem.ResultsA procedure for the site-specific chemical conjugation of hFasLECD with a fluorochrome and functional proteins was devised using an inverse-electron-demand Diels-Alder reaction between trans-cyclooctene group and methyltetrazine group. The conjugations in the present study were attained by using much less molar excess amounts of the compounds to be attached as compared with the conventional chemical modification reactions using maleimide derivatives in the previous study. The isolated conjugates of hFasLECD with sulfo-Cy3, avidin and rabbit IgG Fab’ domain presented the functional and the structural integrities of the attached molecules without impairing the specific binding activity toward human Fas receptor extracellular domain.ConclusionsThe present study provided a new fundamental strategy for the production of the engineered hFasLECDs with additional beneficial functions, which will lead to the developments of the improved diagnostic systems and the effective treatment methods of serious diseases by using this protein as a component of novel molecular tools.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-017-0381-2) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Site-specific chemical conjugation of human Fas ligand extracellular domain using trans-cyclooctene – methyltetrazine reactions

Muraki

Hirota

2017

BMC Biotechnol

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“…A variety of methods, including fluorescence activated cell sorter (FACS) and radioimmunoassay, have been used to measure cell surface antigen expression. Cell ELISA was compared with immunofluoresence staining by Grunow et al ,23 and both assays were found to be similarly sensitive. CD4 positive and CD4 negative cell lines were mixed at different cell ratios.…”

Section: Discussionmentioning

confidence: 99%

“…Using immunoperoxidase cell enzyme linked immunosorbent assay (ELISA),23 24 we quantitatively examined the expression of Fas antigen in salivary ductal cells stimulated by cytokines. A human salivary gland duct cell line (HSG) was established from an irradiated human submandibular salivary gland 25.…”

mentioning

confidence: 99%

Glandular and extraglandular expression of costimulatory molecules in patients with Sjogren's syndrome

Matsumura

Umemiya

Goto

et al. 2001

Annals of the Rheumatic Diseases

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“…Media from hybridoma lines were screened against several cell types by cyto-ELISA (Grunow et al , 1994), flow cytometry and western blot. The monoclonal antibodies produced by the hybridoma lines were isolated using protein G. The target antigens were isolated from cultured hMSC cell homogenate by immunoprecipitation with the purified antibody covalently coupled to the surface of Dynabeads (M270, Invitrogen, Carlsbad, CA, USA) or using Protein G resin (Sigma-Aldrich, Ayshire, UK).…”

Section: Methodsmentioning

confidence: 99%

Tenascin-C and fibronectin expression divide early stage tongue cancer into low- and high-risk groups

et al. 2017

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Background:Oral tongue squamous cell carcinoma (OTSCC) metastasises early, especially to regional lymph nodes. There is an ongoing debate on which early stage (T1-T2N0) patients should be treated with elective neck dissection. We need prognosticators for early stage tongue cancer.Methods:Mice immunisation with human mesenchymal stromal cells resulted in production of antibodies against tenascin-C (TNC) and fibronectin (FN), which were used to stain 178 (98 early stage), oral tongue squamous cell carcinoma samples. Tenascin-C and FN expression in the stroma (negative, moderate or abundant) and tumour cells (negative or positive) were assessed. Similar staining was obtained using corresponding commercial antibodies.Results:Expression of TNC and FN in the stroma, but not in the tumour cells, proved to be excellent prognosticators both in all stages and in early stage cases. Among early stages, when stromal TNC was negative, the 5-year survival rate was 88%. Correspondingly, when FN was negative, no cancer deaths were observed. Five-year survival rates for abundant expression of TNC and FN were 43% and 25%, respectively.Conclusions:Stromal TNC and, especially, FN expressions differentiate patients into low- and high-risk groups. Surgery alone of early stage primary tumours might be adequate when stromal FN is negative. Aggressive treatments should be considered when both TNC and FN are abundant.

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A cell surface ELISA for the screening of monoclonal antibodies to antigens on viable cells in suspension

Cited by 24 publications

References 15 publications

Site-specific chemical conjugation of human Fas ligand extracellular domain using trans-cyclooctene – methyltetrazine reactions

Site-specific chemical conjugation of human Fas ligand extracellular domain using trans-cyclooctene – methyltetrazine reactions

Glandular and extraglandular expression of costimulatory molecules in patients with Sjogren's syndrome

Tenascin-C and fibronectin expression divide early stage tongue cancer into low- and high-risk groups

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