title: Modification of protein with BGL06, a novel branched oligoglycerol derivative.
abstract: A branched oligoglycerol derivative, BGL06, with a cascade-like structure of glycerol units was used as a novel reagent for protein modification. Modification reaction with a recombinant human granulocyte-colony stimulating factor derivative, ND28, was carried out successfully in aqueous conditions to obtain a coupled form, BGL06-ND28. Characterization of the modified ND28 suggests that two types of products were obtained by controlling the reaction; one was H(BGL06)-ND28, a highly modified version coupled with 4.34 molecules of BGL06 units on average, and the other was L(BGL06)-ND28, a moderately modified version coupled with 2.58 molecules of BGL06 units on average, respectively. The properties of these products were compared to the known polyethylene glycol (PEG)-modified ND28. In the cell proliferation assays, unlike PEGylation, modification with BGL06 did not produce a significant loss of biological activity even when the modification extent was elevated. Under such conditions, 76.0% of the activity was in fact maintained for H(BGL06)-ND28, while PEGylated ND28 retained only 24.6% of biological activity in vitro even though the extent of modification was smaller. In addition, H(BGL06)-ND28 showed comparable thermostability to a 20 kDa PEG-modified counterpart. Therefore, the BGL06 derivative will be a useful alternative as a protein modification reagent where PEGylation is not effective.
authors.affiliation: Products Strategy Department, Kyowa Hakko Kogyo Co., Ltd., 1-6-1, Ohtemachi, Chiyoda-ku, Tokyo 100-8185, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Yamaguchi
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Suzawa
authors.forename: Takaaki
authors.initials: T
authors.lastname: Araki
authors.forename: Masaki
authors.initials: M
authors.lastname: Kamiya
authors.forename: Hisao
authors.initials: H
authors.lastname: Nemoto
authors.forename: Motoo
authors.initials: M
authors.lastname: Yamasaki
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D005989:Glycerides / Q000737:chemistry*; D016179:Granulocyte Colony-Stimulating Factor / Q000737:chemistry* / Q000494:pharmacology; D006358:Hot Temperature; D006801:Humans; D011994:Recombinant Proteins; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013388:Succinimides / Q000737:chemistry*
chemical_list: D005989:Glycerides; D011994:Recombinant Proteins; C529082:SuO-BGL06 compound; D013388:Succinimides; D016179:Granulocyte Colony-Stimulating Factor