Presence of bacterial proteins in Human Breast Milk detected through Western Blot Techniques

Breastmilk has always contained a diverse microbiome known as the human milk microbiota (HMM), containing many diverse populations such as bacteria, fungi, parasites, etc. Western blotting is a technique commonly implemented to assist in the detection of antigens or proteins in a sample of interest. The antibodies NTHi 3B9, JLA20, and 15F12F8 are specific for antigens commonly found in bacterial strains including Escherichia coli, Streptococcus pyogenes and staph bacteria. However, the presence of these bacterial proteins in breast milk and evidence behind it are understudied. In order to validate the presence of the specific bacterial proteins in our breast milk samples, we deploy the use of antibodies (NTHi 3B9, JLA20, and 15F12F8) specific to them using the western blot technique. We can report that our bacterial proteins were not detected in control samples using Western Blotting. Consequently, these results show that our antibodies could not be validated for use in breastmilk samples. It is possible that the bacterial proteins were not detected because of the protein concentration of control samples that was added. Future studies should focus on optimizing the protein concentration of samples utilized and overall better precision.