Establishment and Confirmation of a Method for Detection of Total Anti-capsid of AAV8 Antibodies in Serum of Rhesus Monkeys
With the development of gene therapy based on adeno-associated virus(AAV),an important task is to develop a simple,rapid and reliable method for detecting pre-existing anti-AAV antibodies.In this study,a simple and rapid method for the detection of total antibodies(TAb)against AAV8 capsid in serum of rhesus monkeys was developed and confirmed.This method based on enzyme-linked immunosorbent assay(ELISA),and AAV8 capsid was coated on 96-well plate,recombinant protein A/G was used as the detection antibody.The results showed that the multiplicative correction factor of this assay was 1.08 and confirmatory cut point was 29.5％.The screening and confirmatory sensitivities were 43.1 and 76.5 ng/mL,respectively.The positive controls of 1 000,100,50 ng/mL were tolerant to 1.25× 1010,3.13× 109,1.56×109 GC/mL AAV8,respectively.Both the coefficients of variation(CV)of intra-run and inter-run met CV≤20.0％.In addition,the method has sufficient selectivity and sample stability,which can be used for TAb detection of pre-existing anti-AAV8 capsid in serum of rhesus monkeys.
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