Properties of univalent fragments of rabbit antibody isolated by specific adsorption

Nisonoff, Alfred; Wissler, F.C.; Woernley, D.L.

doi:10.1016/0003-9861(60)90229-0

Cited by 79 publications

(20 citation statements)

References 18 publications

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“…The resultant IgG Fc depleted of IgG4 Fc was then used in Western blotting in addition to intact IgG Fc. IgG4 Fc and IgG4 F(ab') 2 were generated through papain [31] and pepsin treatments [32] of purified IgG4 (extracted from pooled patient sera by affinity chromatography as described above) respectively. The step-by-step methodology is provided in Figure S1.…”

Section: Methodsmentioning

confidence: 99%

A Novel Immunoglobulin-Immunoglobulin Interaction in Autoimmunity

et al. 2008

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Well over six decades since its first description, the Rheumatoid Factor (RF)—autoantibodies recognizing Fc (constant) portion of IgG through their own Fab (antigen binding variable segments)—is believed to have come of age. Autoimmune pancreatitis is a unique form of pancreatitis, biologically characterized by an elevated serum IgG4 concentration. Given the fact that IgG4 myeloma proteins can act as RF, we initially hypothesized that IgG4 in autoimmune pancreatitis might do likewise, hence potentially contributing to disease pathogenesis. Indeed Western blotting clearly showed that IgG4 binds to IgG1 κ, IgG2 κ, IgG3 κ myeloma proteins, as well as to IgG Fc, in line with a typical RF activity. Further experiments however unraveled the unexpected fact that unlike hitherto known RF, IgG4 does not engage IgG Fc through its Fab, but its very own Fc. These data therefore collectively describe a Novel RF (NRF) in autoimmune pancreatitis. In the future, the relevance of NRF, beyond autoimmune pancreatitis, in both diagnosis/prognosis as well as pathophysiology of autoimmune and other systemic diseases where IgG4's role seems paramount, needs to be systematically assessed.

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Section: Methodsmentioning

confidence: 99%

A Novel Immunoglobulin-Immunoglobulin Interaction in Autoimmunity

et al. 2008

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“…Purity of the antibody was confirmed by isoelectric focusing (Awdeh et al, 1968) and by polyacrylamide gel electrophoresis (Laemmli, 1976). F(ab')2 fragments were produced by pepsin digestion (Nisonoff et al, 1960) as described elsewhere (Kalofonos et al, 1988a). 4C4 The monoclonal antibody 4C4 (Boniolo et al, 1982) is a mouse IgGI directed against the hepatitis B surface antigen.…”

Section: Patientsmentioning

confidence: 99%

Kinetics, quantitative analysis and radioimmunolocalization using indium-111-HMFG1 monoclonal antibody in patients with breast cancer

Kalofonos

Sackier²,

Hatzistylianou³

et al. 1989

Br J Cancer

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Summary HMFGI tumour associated monoclonal antibody IgGl and F(ab')2 fragments were radiolabelled with indium-111 and used to study patients with breast cancer. In vitro and in vivo stability of the radiolabelled antibodies was shown to be satisfactory. Thirty patients with primary breast cancer underwent tumour resection and quantitative evaluation of the radioactivity in the tumour and normal tissues following administration of specific and non-specific antibodies. The mean tumour uptake of HMFG1 F(ab')2 fragments at 24h was significantly higher (P<0.05) than the intact IgG but at 48h there was no difference. The mean tumour uptake with the specific antibody was higher than the non-specific antibody of the same subclass (P<0.05). Lymph node metastases showed higher antibody uptake than the corresponding primary tumours (P<0.05). Fifteen patients with primary or metastatic breast cancer were investigated by external body scintigraphy using HMFG1 F(ab')2 fragments. Successful localisation was observed in approximately 50% of the primary and metastatic lesions with no false positive results. All the patients had observable concentration of "'1In in the liver (20% of the injected dose), the kidneys and the spleen. Following i.v. administration, F(ab')2 fragments cleared from the blood more rapidly than the intact IgG. We conclude that HMFG1 F(ab')2 fragments can localise specifically and faster than intact IgG in breast cancer but the sensitivity of the radioimmunoscintigraphy is relatively low. This method needs further improvement before becoming clinically useful for detecting and staging breast cancer.

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“…After digestion of rabbit 7S y-globulin with papain at pH 7.4 in the presence of cysteine, Porter [28] separated three fragments by chro matography. Fragments I and II were demonstrated to be univalent by determination of maximal hapten-binding capacity [25]. Nisonoff et al [26] showed that digestion of rabbit 7S y-globulins and anti bodies with pepsin results in a fragment with a sedimentation coef ficient of approximately 5S.…”

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confidence: 99%