Monoclonal antibody selection for interleukin-4 quantification using suspension arrays and forward-phase protein microarrays.
A recombinant mouse interleukin-4 (IL-4) and three different purified rat antimouse IL-4 monoclonal antibodies (Mab) with different clonalities were employed as a model system. This system was used to examine monoclonal antibody effectiveness using both conventional and high-throughput measurement techniques to select antibodies for attaining the most sensitive detection of the recombinant IL-4 through the "sandwich-type" immunoassays. Surface plasmon resonance (SPR) measurements and two high-throughput methods, suspension arrays (also called multiplexed bead arrays) and forward-phase protein microarrays, predicted the same capture (BVD4-1D11) and detection (BVD6-24G2) antibody pair for the most sensitive detection of the recombinant cytokine. By using this antibody pair, we were able to detect as low as 2 pg/mL of IL-4 in buffer solution and 13.5 pg/mL of IL-4 spiked in 100% normal mouse serum with the multiplexed bead arrays. Due to the large amount of material required for SPR measurements, the study suggests that the multiplexed bead arrays and protein microarrays are both suited for the selection of numerous antibodies against the same analyte of interest to meet the need in the areas of systems biology and reproducible clinical diagnostics for better patient care.
Wang
L
L
Biochemical Science Division, National Institute of Standards and Technology, Gaithersburg, Maryland 20899-8312, USA. lili.wang@nist.gov
Cole
K D
KD
Peterson
A
A
He
Hua-Jun
HJ
Gaigalas
A K
AK
Zong
Y
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000918:Antibody Specificity; D001666:Binding Sites, Antibody; D015847:Interleukin-4 / Q000032:analysis* / Q000276:immunology; D051379:Mice; D040081:Protein Array Analysis; D011485:Protein Binding / Q000502:physiology; D051381:Rats; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D015847:Interleukin-4