Detection of anti-tetanus toxoid antibody on modified polyacrylonitrile fibers

Accurate determination of concentration of immunoglobulin (IgG) to tetanus toxoid is important in order to evaluate the immunogenicity of tetanus toxoid vaccines, immune competence in individual patients and to measure the prevalence of immunity in populations. Surface modified polyacrylonitrile (PA...

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Bibliographic Details
Published in:Talanta (Oxford) 2010-10, Vol.82 (5), p.1876-1883
Main Authors: Jain, Swati, Chattopadhyay, Sruti, Jackeray, Richa, Zainul Abid, C.K.V., Kumar, Manoj, Singh, Harpal
Format: Article
Language:English
Subjects:
Acrylic Resins - chemistry
Age
Analytical chemistry
Animals
Antibodies, Bacterial - blood
Chemistry
Enzyme-Linked Immunosorbent Assay - instrumentation
Enzyme-Linked Immunosorbent Assay - methods
Enzymes, Immobilized - chemistry
Exact sciences and technology
Horseradish Peroxidase - chemistry
Humans
Immunoglobulin G - blood
Immunoglobulin G - chemistry
Microscopy, Electron, Scanning
Miscellaneous
Polyacrylonitrile fiber
Rabbits
Reproducibility of Results
Sandwich enzyme linked immunosorbent assay
Sensitivity
Surface Properties
Tetanus toxoid
Tetanus Toxoid - chemistry
Tetanus Toxoid - immunology
Toxoid immobilization
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Summary:Accurate determination of concentration of immunoglobulin (IgG) to tetanus toxoid is important in order to evaluate the immunogenicity of tetanus toxoid vaccines, immune competence in individual patients and to measure the prevalence of immunity in populations. Surface modified polyacrylonitrile (PAN) fibers were evaluated as a matrix to develop highly sensitive method for the detection of anti-tetanus antibody in a sandwich ELISA format. In the proposed method tetanus toxoid immobilized on modified PAN fibers was used to detect anti-tetanus antibody (raised in horse hence represented as horse anti-tetanus toxoid or HAT-Ab) with horse raddish peroxidase enzyme conjugated with Rabbit anti-Horse IgG (RAH-HRP) as the label within 2.5 h. A sigmoidal pattern for the detection of different concentration of antibody ranging from 1.0 to 0.0001 IU mL −1 was validated. The immunoassay recorded a very high sensitivity as concentration as low as 0.0005 IU mL −1 of HAT-Ab was detected. The intra- and inter-assay precision for 3 parallel measurements of 0.01 and for 0.001 IU mL −1 of antibody varied from 5.4% to 11% and 5.7% to 20% respectively. PAN fibers were also used to qualitatively access the presence of different level of anti-tetanus antibody spiked in human blood. Seroepidemiological studies to measure the immunity against tetanus were conducted with twenty-five human beings belonging to various age groups using modified PAN-ELISA. The sensitivity, specificity and the reproducibility of the developed immunoassay indicate the potential application of modified PAN fibers in the field of immunodiagnostics.
ISSN:0039-9140
1873-3573
DOI:10.1016/j.talanta.2010.08.003