Objective To establish and verify the pharmacokineic detection method of porcine anil-human T cell immunoglobulin (p-ATG) , providing technical service for the measurement of total serum p-ATG concentration in patients after clinical medication. Methods Using rabbit anti-pig IgG F(ab')₂ as coating antibody, horseradish peroxidase-labeled goat anti-pig IgG as secondary antibody, a double-antibody sandwich ELISA for p-ATG detection in human serum was established. The standard curve fitting range, parallelism, precision, accuracy and specificity of the method were verified. Using the established method, clinical blood samples of 13 patients administered with different p-ATG doses were tested and the pharmacokinetics was analyzed. Results Starting at 0.44 μg/ml initial concentration, 2¹-2⁷ times diluted standards concentrations and absorbance values showed an S curve with coefficient of determination (R²)>0.99 and recoveries in 89.91%-112.96%. The absolute value of coefficient of variation (CV) between parallelism sample slope and standard slope was<10%, R²>0.99 and CV of R² was<5%. The inter-plate and intra-plate curves CVs were<10%. The repeatability CV was<10% and recoveries were 94.23%-101.80%. The spiked recoveries were 99.87%-104.94%. There were statistically significant differences between sera p-ATG concentrations of post-p-ATG-infused patients and those of healthy and pre-infused individuals (F=1.48, P<0.05). The clinical blood samples from 20, 25 and 30 mg dosage groups showed that the p-ATG half-lives were 20.6, 16.6 and 16.2 d, respectively. Conclusion  The established method has good repeatability, accuracy and specificity, and can be used for the detection of serum concentration of p-ATG after clinical administration.