Diabetes and other autoimmune endocrine diseases
Diabesity, a term for diabetes occurring in the context of obesity, continues to be one of the major human health concerns due to its global rising prevalence. Etiology of diabesity involves insulin resistance, hyperinsulinemia, metabolic dysregulation, and tissue inflammation. Simultaneous quantification of the mediators involved in glucose utilization, adipose tissue growth and development, hormonal control of homeostasis, and immune regulation is required to study the complicated interactions among complex biological processes associated with the metabolic control, homeostasis and development of diabesity. Existing methodologies for detection of metabolic protein regulators require expensive reagents and dedicated instrument. We developed inexpensive bead-based multiplex assay panels for simultaneous quantification of these metabolic mediators using regular flow cytometers commonly available in most biological research and clinical laboratories. The robust quantification panel uses highly specific antibodies and is validated to detect PAI-1, GLP-1 (Total), Insulin, C-peptide, TNF-α, Glucagon, Leptin, Cortisol, IL-1β, IL-6, and GLP-1 (Active) simultaneously in biological samples (serum, plasma, supernatant of cultured cells) with acceptable assay sensitivities, analytical accuracy and reproducibility. Pre- and post-feeding samples collected from normal and diabetic subjects showed expected profile changes for the target proteins. The multiplex assays are customizable for random combinations of targets within the panel and offer an economic and useful tool for obesity and diabetes researchers.