ELISA and multiplex technologies for cytokine measurement in inflammation and aging research.

Over the last decade there has been an enormous expansion of research focused on defining the role of inflammation in aging, age-related diseases, disability, and frailty. The availability of methods to measure cytokines and other inflammatory mediators or markers with high sensitivity and specificity is critically important. Enzyme-linked immunosorbent assay (ELISA), the most widely used and best validated method, is limited by its ability to measure only a single protein in each sample. Recent developments in serum cytokine quantification technology include multiplex arrays, which offer the potential of better evaluating the complexity and dynamic nature of inflammatory responses and offer substantial cost and sample savings over traditional ELISA measurements. Despite potential advantages of this new technology, experience with these techniques is limited, and it has not emerged to date as the gold standard in inflammatory mediator measurement. This article reviews ELISA and the emerging multiplex technologies, compares the cost and effectiveness of recently developed multiplex arrays with traditional ELISA technology, and provides specific recommendations for investigators interested in measuring serum inflammatory mediators in older adults.

[1]  T. Hirano,et al.  Identification of alpha 2-macroglobulin as a carrier protein for IL-6. , 1989, Journal of immunology.

[2]  D. Wakelin,et al.  The Immune Response , 1983 .

[3]  John R. Crowther,et al.  The ELISA Guidebook , 2000, Methods in Molecular Biology™.

[4]  W. Ershler,et al.  Interleukin‐6: A Cytokine for Gerontolgists , 1993, Journal of the American Geriatrics Society.

[5]  J. P. McCoy,et al.  Multiplex bead array assays: performance evaluation and comparison of sensitivity to ELISA. , 2006, Methods.

[6]  U. Prabhakar,et al.  Validation and comparative analysis of a multiplexed assay for the simultaneous quantitative measurement of Th1/Th2 cytokines in human serum and human peripheral blood mononuclear cell culture supernatants. , 2004, Journal of immunological methods.

[7]  J. P. McCoy,et al.  Multiplex bead array assays for detection of soluble cytokines: Comparisons of sensitivity and quantitative values among kits from multiple manufacturers , 2004, Cytometry. Part B, Clinical cytometry.

[8]  J. Ladenson,et al.  Serum versus heparinized plasma for eighteen common chemistry tests: is serum the appropriate specimen? , 1974, American journal of clinical pathology.

[9]  E. Marcantonio,et al.  Chemokines are associated with delirium after cardiac surgery. , 2008, The journals of gerontology. Series A, Biological sciences and medical sciences.

[10]  I. Bautmans,et al.  Heat shock proteins and chemokine/cytokine secretion profile in ageing and inflammation , 2007, Mechanisms of Ageing and Development.

[11]  Fernando A Bozza,et al.  Cytokine profiles as markers of disease severity in sepsis: a multiplex analysis , 2007, Critical care.

[12]  R. Detels,et al.  Variables That Affect Assays for Plasma Cytokines and Soluble Activation Markers , 1999, Clinical Diagnostic Laboratory Immunology.

[13]  S. L. Gonias,et al.  Interleukin-4 and IL-10 bind covalently to activated human alpha2-macroglobulin by a mechanism that requires Cys949. , 2000, Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research.

[14]  S. Berson,et al.  Assay of Plasma Insulin in Human Subjects by Immunological Methods , 1959, Nature.

[15]  Carl Nathan,et al.  Points of control in inflammation , 2002, Nature.

[16]  S. Bandinelli,et al.  A flame burning within , 2004, Aging clinical and experimental research.

[17]  T. Mak,et al.  17 – Cytokines and Cytokine Receptors , 2006 .

[18]  G. Castellani,et al.  Inflammaging and anti-inflammaging: A systemic perspective on aging and longevity emerged from studies in humans , 2007, Mechanisms of Ageing and Development.

[19]  S. Gambino,et al.  A comparison of serum versus heparinized plasma for routine chemistry tests. , 1974, American journal of clinical pathology.

[20]  L. Fried,et al.  Inflammation and Frailty in Older Women , 2007, Journal of the American Geriatrics Society.

[21]  L. Ferrucci,et al.  Inflammation, hormones, and body composition at a crossroad. , 2003, The American journal of medicine.

[22]  L. Ferrucci,et al.  Interleukin-6 in aging and chronic disease: a magnificent pathway. , 2006, The journals of gerontology. Series A, Biological sciences and medical sciences.

[23]  Roger L. Lundblad,et al.  Considerations for the Use of Blood Plasma and Serum for Proteomic Analysis , 2003 .

[24]  A. Hamblin Cytokines and cytokine receptors , 1993 .

[25]  E. Mariani,et al.  Simultaneous evaluation of circulating chemokine and cytokine profiles in elderly subjects by multiplex technology: relationship with zinc status , 2006, Biogerontology.

[26]  S. L. Gonias,et al.  Classification of alpha 2-macroglobulin-cytokine interactions based on affinity of noncovalent association in solution under apparent equilibrium conditions. , 1994, The Journal of biological chemistry.