Optimization, comparison, and application of colorimetric vs. chemiluminescence based indirect sandwich ELISA for measurement of human IL-23.
Currently, there is neither a published ELISA method nor it is clear whether chemiluminescence substrates would provide better sensitivity vs. colorimetric substrates for measuring human IL-23-a recently described Type-1 immunity associated cytokine. Initially, we optimized a colorimetric ELISA using p-nitro-phenyl phosphate substrate. Subsequently, we compared it with chemiluminescence substrates that provided approximately 5-fold enhanced sensitivity (mean sensitivity; 26.3 pg/mL vs. colorimetric assay, 131 pg/mL; p < 0.01). Both methods were reliable, with <10% inter- and intra-assay variations. We then found that the chemiluminescence method was useful in situations where human IL-23 was not readily measurable by a colorimetric method.
Samineni
Sridhar
S
Food Allergy & Immunology Laboratory, Department of Food Science & Human Nutrition, Nutritional Immunology Program, Michigan State University, East Lansing, Michigan 48824, USA
Parvataneni
Sitaram
S
Kelly
Caleb
C
Gangur
Venu
V
Karmaus
Wilfried
W
Brooks
Kevin
K
D000328:Adult; D003124:Colorimetry; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000706:statistics & numerical data; D006801:Humans; D053759:Interleukin-23; D053760:Interleukin-23 Subunit p19; D007378:Interleukins / Q000032:analysis* / Q000097:blood; D049449:Luminescence; D011994:Recombinant Proteins / Q000032:analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
C508840:IL23A protein, human; D053759:Interleukin-23; D053760:Interleukin-23 Subunit p19; D007378:Interleukins; D011994:Recombinant Proteins