Standardizing the Quantification of Total Human Immunoglobulin (IgA and IgG) in Adult Individuals with Autism Spectrum Disorder (ASD)

Autism spectrum disorder (ASD) is a neurodevelopmental disorder that not only affects communication and behavior with often co-occurring gastrointestinal (GI) issues such as constipation and diarrhea. Recent studies have shown that many GI and behavioral symptoms in individuals with ASD are linked to dysregulated immune systems and altered gut microbiomes (bacteria and fungi). In fungal microbiota, a common GI commensal and opportunistic pathogen, Candida, has been found in higher abundance in children with ASD. Few studies have investigated total IgA and IgG levels in both blood and feces of ASD individuals with relatively mixed findings, showing either significantly higher or lower IgG and IgA abundance in ASD vs. TD (typically developing) individuals. Mixed results are likely due to a lack of a standardized method of immunoglobulin (Ig) quantification. In this study, we attempt to standardize an enzyme-linked immunoassay (ELISA) procedure to measure total IgA, total IgG, and anti-Candida albicans IgA and IgG levels in fecal samples of adults with ASD. Measuring Ig levels can reflect altered gut microbiota, GI tract, and immune status in ASD and potentially characterize Ig as a biomarker for ASD. Although we were unable to successfully standardize an Ig ELISA quantification method, SDS-PAGE confirmed the presence of IgA in fecal Ig extracts. Based on our ELISA results, we suspect that dilution factors of fecal Ig extracts need to be modified further to detect the IgA within the detection range. The experimental methodology in this study can be used as a reference to develop and improve a full-proof method of quantifying immunoglobulin from ASD fecal samples, which will help to reveal immune status in ASD.