Challenges and opportunities in fractionation of recombinant human mannan-binding lectin.
Mannan-binding lectin is an oligomeric protein of the innate immune system. Both natural MBL as well as recombinant protein is a heterogeneous composition of oligomers containing from two up to at least eight structural units. Recombinant synthesis in mammalian cell lines often leads to a composition containing small oligomers not abundant in MBL from natural sources such as human plasma. Here, two procedures for fractionation of MBL oligomers are described. In one case, affinity chromatography is employed to enrich the abundance of large MBL oligomers to produce a composition of oligomers resembling natural MBL. In a second case, large-volume ion exchange chromatography is employed to separate these oligomers into fractions with an enhanced size homogeneity. Finally, a well-established procedure for estimating the MBL protein concentration is detailed.
Biophysical Immunology Laboratory, Department of Biomedicine, The Lundbeck Foundation Nanomedicine Center for Individualized Management of Tissue Damage and Regeneration, and Interdisciplinary Nanoscience Center, Aarhus University, Aarhus, Denmark.
Thomas
T
Vorup-Jensen
D005591:Chemical Fractionation / Q000379:methods*; D002846:Chromatography, Affinity / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D006801:Humans; D037601:Mannose-Binding Lectin / Q000737:chemistry / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification*
D037601:Mannose-Binding Lectin; D011994:Recombinant Proteins