NRicher™ C

Enrichment of Complement Cascade Related Proteins

NRicher™ C用于补体级联相关蛋白的富集，Biotech Support Group提供两个规格NRicher™ C，10 preps（NRCO-10）和50 preps（NRCO-50）。

显著特点

· 消耗性的化学衍生磁珠，与物种无关，因为它们不是来自抗体

· 从动物和人的血清/血浆或细胞裂解液中富集补体级联相关蛋白，白蛋白去除率>90%

· 不需要任何专门的仪器，只需要一个微量离心机

· 磁珠格式适合自动化兼容（请咨询）

· 用于LC-MS分析的On-Bead消化，或用于任何功能性、酶促或免疫测定分析的选择性洗脱

The complement cascade is a major component of the immune system that provides powerful host surveillance and protection from invading microbes. It also exerts an important influence on the adaptive immune response by acting synergistically with antibodies as well as promoting B- and T-cell stimulation. The intracellularly active complement system—the complosome, has been shown to play a critical function in regulating T cell responses, cell physiology (such as metabolism), and inflammatory disease processes. Complement dysregulation is implicated in chronic diseases ranging from age-related macular degeneration to neurological & cardiovascular disease, as well as cancer. In acute inflammatory conditions, hyperactive complement may predispose individuals to adverse outcomes, as suspected in hospitalized Covid-19 patients.

The complement system consists of over 50 circulating and membrane proteins, comprising about 5% of the total protein mass in plasma. Most complement proteins circulate in blood as inactive precursors (zymogens), that when triggered, become activated through proteolytic cascades. Although textbooks describe three activation pathways (classical, lectin & alternative), largely under-appreciated is Complement’s evolutionarily conserved link to coagulation to eliminate damaged tissues. Complement can activate platelets and contribute to hemostasis in response to injury. Conversely, complement can be activated from proteolytic enzymes derived from coagulation and fibrinolysis that can cleave both C3 and C5 leading to amplification, and final function. As a result of complement activation, many outcomes are produced including: opsonization of pathogens or damaged-self cells to enhance phagocytosis; production of anaphylatoxins C3a & C5a; recruitment of leukocytes to the inflammatory site; and the terminating end of the cascade – assembly of the membrane attack complex (MAC) on the cell surface.

The terminating end of the complement cascade is derived from the C3 Convertase proteolytic product - C5b, which engages the sequential recruitment of C6, C7, C8, and C9, assembling the membrane attack complex (MAC). Also known as the “terminal complement complex”, it results from the coordination of C5b-7 insertion in the membrane, which then captures C8, inducing polymerization of a C9 ring – to as many as 18, C9’s per pore. Terminal MACs perforate the cell membrane of the invading pathogen or target cell, and when a sufficient number of MAC pores form, the cell dies by osmotic lysis. Sub-lytic doses of MAC however, induce dramatically different effects than lytic doses, including adherence, aggregation, chemotaxis, cell division, and extracellular vesicle release.

In such a critical juncture, the complement system must maintain a delicate balance between activation and inhibition to allow activation when necessary to counteract infectious agents or modified self/host tissues, while concurrently protecting healthy self/host tissue. This protection is achieved systemically through the concerted action of regulators and inhibitors ensuring cell and tissue integrity essential for normal and healthy well-being.
 NRicher™ C provides enrichment of all Complement-related proteins so as to interrogate functional features derived from proteolysis (split products) or gene variants. Specific tryptic peptides that report functional regions are especially noteworthy after NRicher™ C enrichment; for example - C3a, C4BPB, CFB (Ba fragment), peptides have approximately >10X total spectral signal compared to neat (not enriched) serum. Multiplex targeted proteomics can therefore investigate a more complete picture of Complement activation and regulation using NRicher™ C, than would be possible with immunoassay measurements.