Non-antigenic and low allergic gelatin produced by specific digestion with an enzyme-coupled matrix.
Porcine gelatin (heat-denatured collagen) was digested with a bioreactor using an enzyme-coupled matrix (ECM) with purified collagenase. The digested gelatin, FreAlagin type R (M.W. range 200-10000 Da), was further purified by an HPLC system depending upon molecular size. The molecular weight range of the purified fractions, FreAlagin type P and type AD, were 200-500 and 2000-10000 Da, respectively, and glycine was the N-terminal amino acid of both types (> or =93%). ECM has the capability of digesting gelatin at a specific point in the sequence before glycine, and it was determined that FreAlagin type P consists of a tri-peptide fraction with the amino acid sequence Gly-X-Y. No types of FreAlagin exhibited any reactivity with gelatin-specific IgG antibody raised in guinea pigs, and they also possessed an extremely low reactivity with gelatin-specific IgE antibody from the sera of patients who had experienced an anaphylactic reaction against gelatin after vaccination or after eating gelatin-containing foods. From these results, it was determined that FreAlagin types R and AD were non-antigenic, low-allergic gelatins. FreAlagin type R, and especially type AD, had strong adsorption-blocking activity comparable to the level of bovine serum albumin, whereas type P and glycine had virtually no adsorption-blocking activity. Therefore, the new types of gelatin, FreAlagin types R and AD, are suitable for pharmaceutical use to avoid gelatin allergy.
Sakai
Y
Y
BS Laboratory Center, Miyagi Chemical Industrial, CO., Ltd., Sendai, Japan.
Yamato
R
R
Onuma
M
M
Kikuta
T
T
Watanabe
M
M
Nakayama
T
T
D000327:Adsorption; D000818:Animals; D000941:Antigens / Q000737:chemistry / Q000276:immunology*; D002417:Cattle; D003094:Collagen / Q000737:chemistry; D017364:Collagenases / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D005780:Gelatin / Q000737:chemistry / Q000276:immunology*; D007074:Immunoglobulin G / Q000096:biosynthesis; D008970:Molecular Weight; D010592:Pharmaceutic Aids / Q000737:chemistry*; D013552:Swine
D000941:Antigens; D007074:Immunoglobulin G; D010592:Pharmaceutic Aids; D005780:Gelatin; D003094:Collagen; D017364:Collagenases