Carnitine Palmitoyltransferase 2, Mitochondrial (CPT2), member of the carnitine shuttle, is involved in the transport of long-chain fatty acids into the mitochondrial compartment.CPT2 is located in the inner mitochondrial membrane, and its deficiency is an inherited metabolic disease.Three forms of CPT2 deficiency have been described: a myopathic form, a severe infantile form and a neonatal form.Acyl Coenzyme A Dehydrogenase, Short/Very Long Chain (ACADVL) is active toward esters of long-chain and very long chain fatty acids, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant mouse CPT2 and recombinant human ACADVL.Briefly, biotin-linked CPT2 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μl were then transferred to ACADVL-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50μl stop solution to the wells and read at 450nm immediately. The binding activity of recombinant mouse CPT2 and recombinant human ACADVL was shown in Figure 1, the EC50?for this effect is 0.08ug/mL.