Amended on 9 June 2018

I have searched for protein components in cells via the Edman degradation method (please see file; HepG2 fucoidan) without using notorious ELISA method.

Since virus rules gene expression (due thanks to Dr. Joel Subach, Rutgers, The State University of New Jersey, New Brunswick, NJ, USA), I have studied about the abnormal expression of Mucin, which is usually expressed in the Epithelial cells.

Healed hepatocyte HepG2 (cultured with fucoidan) has high amount of Mucin-16 at 67.4 μg/mg of cell protein (6.7% among cell proteins). This glycoprotein is a component of the ocular surface (including the cornea and conjunctiva), the respiratory tract, and the female reproductive tract epithelia.

Mucin genes in cultured liver cells seem to be up-regulated by HHVs (except HHV-5 and HHV-6) and down-regulated by HIV and HCV.

HCC tissue (named as No.6) has Mucin 2/Intestinal mucin-2 at 10.6 μg/mg of tissue protein. This protein is present in colon, small intestine, colonic tumors, bronchus, cervix, and gall bladder. This tissue has Herpes simplex virus 1/HHV-1 at 8.2, Herpes simplex virus 2/HHV-2 at 14.6, Varicella-Zoster virus/HHV-3 at 0.25, and Epstein-Barr virus/HHV-4 at 5.4 μg/mg of tissue protein, respectively.

Human breast milk has Mucin-5AC/Gastric mucin at 2.7 μg/mg of milk protein. This glycoprotein is a gel-forming protein of gastric and respiratoy tract epithelia that protects the mucosa from infection and chemical damage by binding to inhaled microrganisms and particles that are subsequently removed by the mucocilary system. Thus this protein in the human breast milk seems to be excreted to protect the baby from viral infection. Mucin is not excreted in bovine milk. This human milk contains Epstein-Barr virus/EBV/Human herpesvirus 4/HHV-4 at 7.7 μg/mg of milk protein and Human herpesvirus 7/HHV-7 at 2.6 μg/mg of milk protein (our unpublished observation).

Serum of GSD-1b patient (3y11mo, female; at 2mo after the liver transplantation) has Mucin-6/Gastric mucin-6 at 10.8 μg/mg of serum protein. This serum uniquely has SAICAR synthetase (Wolbachia sp.) at 6.9 μg/mg of serum protein. Then, excretion of Mucin-6 may be induced by Wolbachia.

Serum of healthy male (33y) has Mucin-5B/Cervical mucin/Sublingual gland mucin at 29.9 μg/mg of serum protein. This person has no bacteria, and he may have been injected Doxycycline. Therefore, estimation of presence of Wolbachia in his serum has become to be not possible.

Notorious Western Blotting/Immunological assay and ELISA method are both non quantitative methods.

Therefore, I recommend the followings; i.e.,

1st) Please separate the samples by reliable HPLC-Surf-SEC method (please see file; SEC column 300 A silica).

2nd) Please fractionate the protein peak, and identify the protein peak by the PDMD method (please see again file; HepG2 fucoidan). Please use a reliable Shimadzu PPSQ protein sequencer.

Please have a look at these useful links.

https://www.researchgate.net/post/Protocol_to_prepare_mice_fecal_sample_for_measuring_IgA_and_Mucin_2_using_ELISA

http://search.cosmobio.co.jp/cosmo_search_p/search_gate2/docs/csr_/ffamuk01e.20141001.pdf