title: Overproduction of recombinant human mannose-binding lectin (MBL) in Chinese hamster ovary cells.
abstract: Mannose-binding lectin (MBL) is an important serum protein that functions in the innate immune system and has been considered to have therapeutic potential in MBL replacement therapies for patients with deficient or low levels of MBL. In this study, we established a Chinese hamster ovary (CHO) cell line that overexpresses the recombinant human MBL (rhMBL) protein. In an 11-day batch culture process using a 30-L bioreactor (20-L working volume) and serum-free medium, these cells could produce over 226 mg/L of rhMBL protein. The recombinant protein was then purified to homogeneity from the culture supernatant using a three-step chromatographic procedure that resulted in a recovery rate of approximately 55%. This purified rhMBL protein adopted oligomeric bouquet-like structures that were similar to those of native MBL present in human blood, and these oligomeric structures were reported to be critical in MBL functions. We further demonstrated in carbohydrate binding and complementation activation assays that this rhMBL protein was functionally active with very similar dissociation constants and half maximal effective concentrations to those of native MBL.
authors.affiliation: R&D Center, CHA Vaccine Institute, Seongnam-si, Republic of Korea.
authors.forename: Byung Cheol
authors.initials: BC
authors.lastname: Ahn
authors.forename: Jeong Soo
authors.initials: JS
authors.lastname: Park
authors.forename: Dongjun
authors.initials: D
authors.lastname: Kim
authors.forename: Junho
authors.initials: J
authors.lastname: Park
authors.forename: Jia
authors.initials: J
authors.lastname: Pi
authors.forename: Jung Sun
authors.initials: JS
authors.lastname: Yum
authors.forename: Yongsu
authors.initials: Y
authors.lastname: Jeong
authors.forename: Kwanghee
authors.initials: K
authors.lastname: Baek
authors.forename: Hong Mo
authors.initials: HM
authors.lastname: Moon
authors.forename: Jaeseung
authors.initials: J
authors.lastname: Yoon
mesh_terms: D000818:Animals; D019149:Bioreactors; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D037601:Mannose-Binding Lectin / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D037601:Mannose-Binding Lectin; D011994:Recombinant Proteins