Production of monoclonal antibodies to human IgM for assay of viral IgM antibodies

Forghani, Bagher; Myoraku, Carolyn K.; Schmidt, Nathalie J.

doi:10.1016/0166-0934(82)90023-4

Cited by 16 publications

(5 citation statements)

References 10 publications

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“…Laboratory B (California Department of Health Services) used an EIA based on a lysate of induced BC3 cells 24 and two BC3 mIFAs, one based on the latent nuclear antigen present in uninduced cells and the other based on cells that had been induced to lytic HHV‐8 replication by treatment with tetradecanoyl phorbol acetate (TPA). A pool of MoAbs against human IgG were used in the mIFA 25 . For the purpose of analysis, indeterminate and nonspecific reactors were treated as seronegative results.…”

Section: Methodsmentioning

confidence: 99%

Multicenter comparison of serologic assays and estimation of human herpesvirus 8 seroprevalence among US blood donors

Pellett

Wright²,

Engels³

et al. 2003

Transfusion

151

141

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Medical and behavioral screening does not eliminate HHV-8-seropositive persons from the US blood donor pool, but no viral DNA was found in donor blood. Further studies of much larger numbers of seropositive individuals will be required to more completely assess the rate of viremia and possibility of HHV-8 transfusion transmission. Current data do not indicate a need to screen US blood donors for HHV-8.

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Section: Methodsmentioning

confidence: 99%

Multicenter comparison of serologic assays and estimation of human herpesvirus 8 seroprevalence among US blood donors

Pellett

Wright²,

Engels³

et al. 2003

Transfusion

151

141

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“…Immune reagents. The production and characterization of a clone of monoclonal antibody to human IgM have been described in detail (7). Antiserum to measles virus was produced by intraperitoneal inoculation of hamsters with suckling hamster brain infected with the neurotropic HNT Philadelphia strain 26 (2) of measles virus.…”

Section: Methodsmentioning

confidence: 99%

“…However, the development and application of viral IgM antibody assays has been hampered by lack of anti-human IgM reagents of adequate specificity, potency, and consistency. We have recently described the production of monoclonal antibodies to human IgM, together with preliminary data on their suitability for assay of viral IgM antibodies (7). In the present report we describe results obtained by using the monoclonal antibodies as "capture" antibodies in a solid phase for assay of IgM immunoglobulins to measles and rubella viruses by enzyme immunofluorescence assays (EIFA).…”

mentioning

confidence: 99%

Use of monoclonal antibodies to human immunoglobulin M in "capture" assays for measles and rubella immunoglobulin M

Forghani¹,

Myoraku²,

Schmidt³

1983

J Clin Microbiol

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Monoclonal antibodies to human immunoglobulin M (IgM) were used in a fourphase enzyme immunofluorescence "capture" assay for determination of IgM antibodies to measles and rubella viruses. Little or no background reactivity was seen in the test system, and interfering effects of rheumatoid factor were avoided by preabsorption of test sera with aggregated human IgG. Virus-specific IgM antibody was demonstrable in 23 of 24 patients with serological evidence of measles virus infections and in 36 of 36 patients with serological evidence of postnatal rubella infection. A few of the rubella patients did not show IgM antibody until 5 days after onset of illness. The enzyme immunofluorescence assay was able to demonstrate rubella IgM antibody in congenitally infected newborns, whereas indirect immunofluorescence results for virus-specific IgM were negative. Viral IgM antibody was not detected in persons with past infections with the test viruses, in young children without evidence of past infection, or in patients infected with heterotypic viruses, rickettsiae, chlamydiae, or mycoplasmas.

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“…This should encourage the development of antibody class capture assays for use in other infectious diseases where IgM detection is of particular value in immunodiagnosis. 19 The anti-T gondii monoclonal antibody C1E3 was selected specifically for its ability to induce complement mediated cell lysis of the living parasite in the methylene blue dye test, regarded as the definitive test system for the detection of antibodies against this organism. This antibody is of the IgG3 subclass.…”

Section: Balfour Harford Goodall Discusionmentioning

confidence: 99%

Use of monoclonal antibodies in an ELISA to detect IgM class antibodies specific for Toxoplasma gondii.

Balfour

Harford²,

Goodall³

1987

J Clin Pathol

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SUMMARY Two monoclonal antibodies CH6 and Cl E3 were used in an antibody class capture assay for the detection of IgM antibodies specific for Toxoplasma gondii. CH6 was used on the solid phase to capture human IgM. After a Toxoplasma gondii antigen had been added, specifically bound material was detected using Cl E3 coupled to horseradish peroxidase. The assay was compared with an established system using polyclonal antisera at both the capture and antigen detection stages. A good correlation was found, with 97 3% (125 of 128) of sera giving the same classification in both assays. Three sera were positive only in the polyclonal system. No false positive results were found when 118 negative sera were examined.The These problems can be overcome by the antibody class capture assay (ACCA).6 Antisera bound to a solid phase is used to capture IgM from the test serum and the remaining serum components washed away. The specificity of the bound IgM can then be assayed by the addition of the antigen of interest, followed by an enzyme labelled antibody against this antigen. Standardisation of both the capture antibody and the enzyme labelled antibody of specific interest is important to ensure that the sensitivity, specificity, and reproducibility of the assay are optimised.Although an antihuman IgM monoclonal antibody Tibi 82 has been used in a radioimmunoassay and in Accepted for publication 9 March 1987 two reverse immunosorbent methods for specific IgM detection, it was not used in an enzyme linked immunosorbent assay (ELISA) system.7 8 In this study an antihuman IgM monoclonal antibody CH6 was used in an ACCA ELISA to detect IgM antibodies specific for Toxoplasma gondii and a second monoclonal antibody, C1E3, used at the antigen detection stage. Cl E3 recognises a major membrane protein antigen of Tgondii and will induce complement mediated cell lysis of the organism. This assay was evaluated and compared with an established ACCA ELISA in which polyclonal antisera were used. Material and methodsSera routinely submitted for toxoplasma serology were titrated in both the dye test and the indirect haemagglutination test (IHAT).9 A total of 128 sera with a dye test titre of > 32 were subsequently tested in the two ACCA enzyme linked immunoabsorbent assay (ELISA) systems for specific IgM. A further 118 sera with no detectable antibody were used to test the specificity of the monoclonal assay.

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Production of monoclonal antibodies to human IgM for assay of viral IgM antibodies

Cited by 16 publications

References 10 publications

Multicenter comparison of serologic assays and estimation of human herpesvirus 8 seroprevalence among US blood donors

Multicenter comparison of serologic assays and estimation of human herpesvirus 8 seroprevalence among US blood donors

Use of monoclonal antibodies to human immunoglobulin M in "capture" assays for measles and rubella immunoglobulin M

Use of monoclonal antibodies in an ELISA to detect IgM class antibodies specific for Toxoplasma gondii.

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