Evaluation of Thirty-One Mouse Monoclonal Antibodies to Human IgG Epitopes

Reimer, Charles B.; Phillips, Donald J.; Aloisio, Carol H.; Moore, Deborah; Galland, G. Gale; Wells, Thomas W.; Black, C M; McDougal, J S

doi:10.1089/hyb.1984.3.263

Cited by 106 publications

(30 citation statements)

References 18 publications

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“…Virtually no cross-reactivity with immunoglobulins of goat, rab bit, horse and sheep was defined. Our results are in agreement with the suggestion that by using McAb, assay specificity, sensitivity, precision and accuracy in the ELISA can be simultaneously improved [18,20,34], Use of a fluorogenic substrate with a time-resolved fluorometer has been suggested to provide a signifi cant advantage in the measurement of antibodies [22][23][24][25]. Our experience with an AP-G used with a flu orogenic substrate indicated that nonspecific binding was consistently low (<7.1% of top standard curve point).…”

Section: Discussionsupporting

confidence: 92%

Measurement of IgG-Blocking Antibodies by ELISA Using Monoclonal Antibody and Fluorogenic Substrate

Lee

Rommel

Adkinson³

1988

Int Arch Allergy Immunol

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An enzyme-linked immunosorbent assay (ELISA) using mouse monoclonal antihuman γ-chain antibody and a fluorogenic substrate has been developed for quantitation of IgG-blocking antibodies in human serum. Generation of fluorescent product was linear with time to 60 min. Using optimal conditions the ELISA was sensitive to < 1 ng/ml of specific IgG to short ragweed pollen. The assay demonstrated consistently parallel dilution curves with 51 sera (mean interdilutional coefficient of variation = 8.8%). Reproducibility was determined by constructing precision profiles for intra and interassay variation for the entire working range of the assay. Intraassay CVs ranged from a mean of 13% at threshold to < 5% at higher antibody concentration. Interassay reproducibility similarly ranged from 18 to 10%. In this assay the effect of serum dilution on nonspecific binding was minimal and specific binding of 4–10 ng IgG antibody to the antigen-adsorbed wells was largely complete (75.8 ± 4.8%) and highly specific ( > 98%). This application of ELISA for ragweed IgG antibody measurement has performance specifications equal or superior to previously developed radioimmunoassay and ELISA systems.

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Section: Discussionsupporting

confidence: 92%

Measurement of IgG-Blocking Antibodies by ELISA Using Monoclonal Antibody and Fluorogenic Substrate

Lee

Rommel

Adkinson³

1988

Int Arch Allergy Immunol

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“…Monoclonal antibodies to human IgG1 (HP6069 ␥1 Fc), IgG2 (HP6002 ␥1 Fc), IgG3 (HP6047, anti-hinge region), and IgG4 (HP6025 ␥1 Fc) (12,34) were purchased from the Hybridoma Reagent Laboratory. For determination of a specific IgG subclass concentration, Immulon 2 HB microtiter plates (Thermo Labsystems) were coated with the appropriate monoclonal anti-subclass capture antibody at a concentration of 2 g/ml in 0.01 M PBS (Life Technologies), pH 7.4, at 4°C for 16 to 24 h. The USNRP was used to generate 7-point standard curves in triplicate in a threefold dilution series (1:100 to 1:12,800).…”

Section: Methodsmentioning

confidence: 99%

Mass Value Assignment of Total and Subclass Immunoglobulin G in a Human Standard Anthrax Reference Serum

Semenova

Steward‐Clark

Stamey

et al. 2004

Clin Vaccine Immunol

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An anti-Anthrax Vaccine Adsorbed (anti-AVA) standard human reference serum pool, AVR414, has been prepared, and the total and protective antigen (PA)-specific immunoglobulin G (IgG) were quantified. AVR414 was prepared by plasmapheresis of healthy adults who had received a minimum of four subcutaneous injections of AVA. Mass values (in milligrams per milliliter) for total IgG and IgG subclasses 1 to 4 were determined by radial immunodiffusion. Anti-PA-specific IgG assignment (in micrograms per milliliter) was done by consensus of two complementary approaches: homologous enzyme-linked immunosorbent assay (ELISA) with affinity-purified anti-PA IgG as a calibrator and summation of mean PA-specific IgG subclass concentrations determined by IgG subclass-specific ELISA using the United States National Reference Preparation for Human Serum Proteins as a standard. The total IgG concentration assigned to AVR414 reference serum was 8.33 mg/ml. IgG subclass concentrations were the following: for IgG1, 4.48 mg/ml; for IgG2, 3.35 mg/ml; for IgG3, 0.37 mg/ml; and for IgG4, 0.30 mg/ml. The assigned mass value for total anti-PA-specific IgG was 141.2 g/ml. Anti-PA-specific IgG subclass concentrations were the following: for IgG1, 79.6 g/ml; for IgG2, 35.3 g/ml; for IgG3, 3.2 g/ml; and for IgG4, 25.3 g/ml. Human reference serum pool AVR414 will have direct application in the standardization of anthrax serological assays, in reagent qualification, and as a standard for quantification of PA-specific IgG in humans who have been vaccinated with or otherwise exposed to Bacillus anthracis PA.The immune response to anthrax toxin protective antigen (PA) is central to protection against anthrax (19,20). Immunoglobulin G (IgG) is the most abundant immunoglobulin in human serum and provides the dominant immune response to protein antigens after vaccination with multiple injections (16, 16a). Measurement of anti-PA IgG antibody is therefore an appropriate marker of human immune responses to Bacillus anthracis infection and anthrax vaccines. A lack of assay standardization and qualified reagents has been a major obstacle to the comparative analysis of human serological responses to clinical anthrax and anthrax vaccines. Compounding this problem are variations in antigen selection, preparation, and purity; variations in assay methodology and end point determination between laboratories; the diversity of antibodies in polyclonal serum; and the absence of a suitable standard reference serum (32). In 2001, the Centers for Disease Control and Prevention (CDC; Atlanta, Ga.) initiated the Anthrax Vaccine Research Program to determine the feasibility of reducing the number of priming series doses of the licensed Anthrax Vaccine Adsorbed (AVA or BioThrax; BioPort Corp., Lansing, Mich.) (17,26,27) from six to three and changing the route of administration from subcutaneous (s.c.) to intramuscular (28) without reducing the vaccine's immunogenicity. The Anthrax Vaccine Research Program required the development of precise, accurate, specific, and sensiti...

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“…These monoclonal antibodies were found to be specific in ELISA. [23][24][25][26] The normal absorbance range for each IgG subclass was obtained by analyzing normal serum from healthy volunteers. Ranges of absorbance (average Ϯ SD) of normal individuals tested at 20-fold plasma dilution for each subclass were 0.071 Ϯ 0.021 for IgG1, 0.068 Ϯ 0.019 for IgG2, 0.073 Ϯ 0.022 for IgG3, and 0.065 Ϯ 0.023 for IgG4.…”

Section: Identification Of Immunoglobulin Class and Subclass Of Anti-mentioning

confidence: 99%

Functional mapping of anti-factor IX inhibitors developed in patients with severe hemophilia B

Olivier

Lenting²,

Cherel³

et al. 2001

Blood

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Development of inhibitory antibodies is a serious complication of treatment with repeated factor IX infusions in a minority of patients with hemophilia B. Such antibodies detected in 8 patients have been characterized. Typing studies revealed that patients' immune response toward factor IX is highly heterogeneous and involves immunoglobulin G (IgG) antibodies, preferentially IgG1 and IgG4. The preservation of the sequence and the 3-dimensional orientation of the amino acids constituting one epitope are highly important for the assembly of an antibodyantigen complex. To localize the epitopes on the factor IX molecule, an original approach was designed using a set of factor X chimeras carrying regions of factor IX. Results showed that some patients' antibodies were directed against both the domain containing the ␥-carboxy glutamic acid residues (Gla domain) and the protease domain of factor IX. In contrast, no binding was observed to the epidermal growth factor-like domains or to the activation peptide. Functional characterization showed that the purified IgG from patients' serum inhibited the factor VIIIa-dependent activation of factor X. Moreover, patients' IgG directed against the Gla domain inhibited the binding of factor IX to phospholipids as well as the binding of factor VIII light chain to factor IXa. These data demonstrate that inhibitors appearing in patients with severe hemophilia B display specificity against restricted functional domains of factor IX. (Blood. 2001;98:1416-1423)

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Evaluation of Thirty-One Mouse Monoclonal Antibodies to Human IgG Epitopes

Cited by 106 publications

References 18 publications

Measurement of IgG-Blocking Antibodies by ELISA Using Monoclonal Antibody and Fluorogenic Substrate

Measurement of IgG-Blocking Antibodies by ELISA Using Monoclonal Antibody and Fluorogenic Substrate

Mass Value Assignment of Total and Subclass Immunoglobulin G in a Human Standard Anthrax Reference Serum

Functional mapping of anti-factor IX inhibitors developed in patients with severe hemophilia B

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