A simple, non-chromatographic purification procedure for monoclonal antibodies Isolation of monoclonal antibodies against cytochrome P450 isozymes

Reik, Linda M.; Maines, Sarah L.; Ryan, Dene E.; Levin, Wayne; Bandiera, Stelvio M.; Thomas, Paul E.

doi:10.1016/0022-1759(87)90180-3

Cited by 164 publications

(45 citation statements)

References 9 publications

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“…Four mAbs (ITEM-1, ITEM-2, ITEM-3, and ITEM-4) were identified by their cytotoxic effects and cloned by limiting dilution. ITEM-1 (mouse IgG1/ ), ITEM-2 (mouse IgA/ ), ITEM-3 (mouse IgG2b/ ), and ITEM-4 (mouse IgG2b/ ) were purified from culture supernatants by caprylic acid and ammonium sulfate precipitation method (19).…”

Section: Generation Of Anti-hfn14 Mabsmentioning

confidence: 99%

Fibroblast Growth Factor-Inducible 14 Mediates Multiple Pathways of TWEAK-Induced Cell Death

Nakayama¹,

Ishidoh²,

Kojima³

et al. 2003

The Journal of Immunology

132

133

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TWEAK, a TNF family member, is produced by IFN-γ-stimulated monocytes and induces multiple pathways of cell death, including caspase-dependent apoptosis, cathepsin B-dependent necrosis, and endogenous TNF-α-mediated cell death, in a cell type-specific manner. However, the TWEAK receptor(s) that mediates these multiple death pathways remains to be identified. Recently, fibroblast growth factor-inducible 14 (Fn14) has been identified to be a TWEAK receptor, which was responsible for TWEAK-induced proliferation of endothelial cells and angiogenesis. Because Fn14 lacks the cytoplasmic death domain, it remains unclear whether Fn14 can also mediate the TWEAK-induced cell death. In this study, we demonstrated that TWEAK could induce apoptotic cell death in Fn14 transfectants. A pan-caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone, rather sensitized the Fn14 transfectants to TWEAK-induced cell death by necrosis via reactive oxygen intermediates and cathepsin B-dependent pathway. By using newly generated agonistic anti-Fn14 mAbs, we also observed that Fn14 is constitutively expressed on the cell surface of all TWEAK-sensitive tumor cell lines, and can transmit the multiple death signals. Moreover, an anti-Fn14 mAb that blocks TWEAK-Fn14 interaction could totally abrogate TWEAK binding and TWEAK-induced cell death in all TWEAK-sensitive tumor cell lines. These results revealed that the multiple pathways of TWEAK-induced cell death are solely mediated by Fn14.

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Section: Generation Of Anti-hfn14 Mabsmentioning

confidence: 99%

Fibroblast Growth Factor-Inducible 14 Mediates Multiple Pathways of TWEAK-Induced Cell Death

Nakayama¹,

Ishidoh²,

Kojima³

et al. 2003

The Journal of Immunology

132

133

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“…Those tested during this study are fully described in Table I. mAbs were purified using either caprylic acid precipitation (39) or protein A affinity chromatography. Epitope mapping for identifying linear epitopes possibly recognized by mAbs was performed as described in Morel et al (40).…”

Section: Methodsmentioning

confidence: 99%

Screening of 145 Anti-PrP Monoclonal Antibodies for Their Capacity to Inhibit PrPSc Replication in Infected Cells

Féraudet

Morel

Simon

et al. 2005

Journal of Biological Chemistry

286

267

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Prion diseases are transmissible neurodegenerative disorders affecting humans and animals for which no therapeutic or prophylactic regimens exist. During the last three years several studies have shown that antiPrP monoclonal antibodies (mAbs) can antagonize prion propagation in vitro and in vivo, but the mechanisms of inhibition are not known so far. To identify the most powerful mAbs and characterize more precisely the therapeutic effect of anti-PrP antibodies, we have screened 145 different mAbs produced in our laboratory for their capacity to cure cells constitutively expressing PrPSc. Our results confirm for a very large series of antibodies that mAbs recognizing cell-surface native PrPc can efficiently clean and definitively cure infected cells. Antibodies having a cleaning effect are directed against linear epitopes located in at least four different regions of PrP, suggesting an epitope-independent inhibition mechanism. The consequence of antibody binding is the sequestration of PrPc at the cell surface, an increase of PrPc levels recovered in cell culture medium, and an internalization of antibodies. Taken together these data suggest that the cleaning process is more likely due to a global effect on the PrP trafficking and/or transconformation process. Two antibodies, Sha31 and BAR236, show an IC 50 of 0.6 nM, thus appearing 10-fold more efficient than previous antibodies described in the literature. Finally, five co-treatments were also tested, and only one of them, described previously (SAF34 ؉ SAF61), lowered PrPSc levels in the cells synergistically.

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“…15,16 They recognized different epitopes along the PrP sequence (Table 2). Monoclonal antibodies were purified using either caprylic acid precipitation 17 or protein A affinity chromatography. Epitope mapping for identifying linear epitopes possibly recognized by monoclonal antibodies was performed as previously described.…”

Section: Anti-prp Antibodiesmentioning

confidence: 99%

Human prion diseases: from antibody screening to a standardized fast immunodiagnosis using automation

et al. 2008

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Demonstration of pathological prion protein accumulation in the central nervous system is required to establish the diagnosis of transmissible subacute encephalopathies. In humans, this is frequently achieved using prion protein immunohistochemistry in paraffin-embedded tissue, a technique that requires multiple epitope retrieval and denaturing pretreatments. In addition to being time-consuming, this procedure induces tissue alterations that preclude accurate morphological examination. The aim of this study was to simplify prion protein immunohistochemistry procedure in human tissue, together with increased sensitivity and specificity. We screened a panel of 50 monoclonal antibodies produced using various immunogens (human and ovine recombinant prion protein, prion protein peptides, denatured scrapie-associated fibrils from 263K-infected Syrian hamsters) and directed against different epitopes along the human prion protein sequence. A panel of different forms of genetic, infectious and sporadic transmissible subacute encephalopathies was assessed. The monoclonal 12F10 antibody provided a high specificity and fast immunodiagnosis with very limited denaturing pretreatments. A standardized and reliable fast immunostaining procedure was established using an automated diagnostic system (Nexes, Ventana Medical Systems) and allowed prion protein detection in the central nervous system and in tonsil biopsies. It was evaluated in a series of 300 patients with a suspected diagnosis of transmissible subacute encephalopathies and showed high sensitivity and specificity.

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A simple, non-chromatographic purification procedure for monoclonal antibodies Isolation of monoclonal antibodies against cytochrome P450 isozymes

Cited by 164 publications

References 9 publications

Fibroblast Growth Factor-Inducible 14 Mediates Multiple Pathways of TWEAK-Induced Cell Death

Fibroblast Growth Factor-Inducible 14 Mediates Multiple Pathways of TWEAK-Induced Cell Death

Screening of 145 Anti-PrP Monoclonal Antibodies for Their Capacity to Inhibit PrPSc Replication in Infected Cells

Human prion diseases: from antibody screening to a standardized fast immunodiagnosis using automation

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