SEC-MALS is a powerful technique that combines multi-angle light scattering with size-exclusion chromatography. The technique enables the absolute characterization of multiple properties of macromolecules, such as molar mass, sizes, conformation, as well as conjugation ratio. With our well-established platforms, the experienced scientists here at Creative Biolabs is dedicated to helping you with different purposes. Here, we briefly describe the C2 SEC MALS protocol to help you characterize complement C2 more easily. Please note that our protocols are only for your reference.
C2 SEC MALS Protocol
Fig.1 Flow chart of C2 SEC MALS protocol. (Creative Biolabs)
Published Data
Fig.2 SEC-MALS evaluation of glycosylation patterns in recombinant human C2 (rhC2).1
Multi-angle static light scattering (MALS) measurements revealed that intact recombinant human C2 (rhC2), including its glycosylation, exhibits a weight-average molar mass (Mw) of 99,800 g/mol. The number-average molar mass (Mn) was determined concurrently by SEC-MALS, yielding a polydispersity index of 1.00054 indicative of a narrow mass distribution. Differential refractive index (dRI) versus elution time confirmed rhC2 elution, with light scattering and 280 nm absorbance establishing a protein core mass of 84,000 g/mol and a glycosylation contribution of 15,800 g/mol. Size exclusion chromatography demonstrated purity greater than 95%, with minor C2b impurities validated by MALDI-TOF analysis as confirmed independently.
To fill some of the gaps in the field of the human complement system, Creative Biolabs now offers a range of complementary products and complement assay services for our clients all over the world. Our service can be tailor-designed to meet your specific needs.
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If you are interested in our products and services, please do not hesitate to contact us for more information.
Reference
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Martini, Paolo GV, et al. "Recombinant human complement component C2 produced in a human cell line restores the classical complement pathway activity in-vitro: an alternative treatment for C2 deficiency diseases." BMC Immunology 11 (2010): 1-10. Distributed under Open Access license CC BY 2.0, without modification.
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