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Annals of Clinical & Laboratory Science 35:312-317 (2005)
© 2005 Association of Clinical Scientists

Microplate ELISAs for Soluble VCAM-1 and ICAM-1

Pi-Yueh Chang1, Tsu-Lan Wu1, Kuo-Chien Tsao1, Chia-Chi Li1, Chien-Feng Sun1 and James T. Wu2
1 Department of Pathology, Chang Gung Memorial Hospital, Taipei, Taiwan2 ARUP Laboratories, Department of Pathology, University of Utah Medical Center, Salt Lake City, Utah

Address correspondence to James T. Wu, Ph.D., ARUP Laboratories, 500 Chipeta Way, Salt Lake City, UT 84108, USA; tel 801 583 2787; fax 801 584 5207; e-mail wuj{at}aruplab.com.

Soluble vascular cell adhesion molecule (sVCAM-1) and soluble intercellular adhesion molecule (sICAM-1) are adhesion molecules that are detectable in the serum of patients with cancer, cardiovascular diseases (CVD), and type 2 diabetes. This report describes enzyme-linked immunosorbent assays (ELISAs) on microplates for sVCAM-1 and sICAM-1. The ELISAs have the sandwich test format; polyclonal antibodies are coated on microwells and a one-step procedure is used in which the serum specimen and detecting antibody are added simultaneously to an antibody-coated well. These assays both use HRP-conjugated sheep anti-mouse-IgG to generate the color for quantification. Sensitivities for detecting sVCAM-1 and sICAM-1 are 49 and 40 ng/ml, respectively. Coefficients of variation for within-day and day-to-day replicate analyses are <10%. Results by these in-house ELISAs for serum sVCAM-1 and sICAM-1 compared well with those obtained with commercial kits from R&D Systems, Inc. (correlation coefficients = 0.98 and 0.99 for sVCAM-1 and sICAM-1, respectively). Reference values for serum sVCAM-1 and sICAM-1 levels were measured in 369 apparently healthy Chinese adults, age 30 to 79 yr. There was no significant effect of gender on the reference values for sVCAM-1 or sICAM-1. Serum sVCAM-1 levels (mean ± SD) were higher in subjects 60 yr old (625 ± 126 ng/ml), compar ed to those <60 yr old (525 ± 110 ng/ml) (p <0.001). Age did not significantly affect the reference values for serum sICAM-1 levels (mean ± SD, 249 ± 86 ng/ml). The authors believe that these simple, inexpensive ELISAs will be useful for assessing the risks for development of cancer, CVD, and type 2 diabetes.

Keywords: sVCAM-1, sICAM-1, adhesion molecules, ELISA, microtiter plate

Abbreviations: BSA. bovine serum albumin; ELISA, enzyme-linked immunosorbent assay; HRP, horse radish peroxidase; sICAM, soluble intercellular adhesion molecule; NF-{kappa}B, nuclear transcription factor-kappa B; PBS, phosphate buffered saline; TMB, tetramethylbenzidine (K-Blue); sVCAM, soluble vascular cell adhesion molecule




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