HYBRIDOMA TECH TO PLASMA REGULATORY PROTEINS FOR COMPLEMENT

hybridoma technology

• formation of a hybridoma in monoclonal antibody production

• a mouse is immunized, and spleen cells (plasma or b cells) are removed

• these cells are fused with non-secreting myeloma cells and thne plated in a restrictive medium

• only the hybridoma cells will grown in this medium, where they synthesize and secrete a monoclonal immunoglobulin specific for a single determinant on an antigen

hybridoma

• an immortal cell capable of producing indefinite sequence of nucleotides

mouse

• animal used for hybridoma

polyethylene glycol

• for fusion of plasma cell with myeloma cell

• surfactant used for hybridoma

medium with hat (hypoxanthine, aminopterin, and thymidine)

• selective medium used for hybridoma

monoclonal antibodies

• purified antibodies cloned from a single cell

• these antibodies exhibit exceptional purity and specificity and are to recognize and bind to a specific antigen

complement system

• a humoral mechanism of nonspecific immune response consisting of 14 distinct serum proteins (nine components) that proceed in a cascading sequence of activation, resulting in cell lysis

anaphylatoxin

• a small peptide that cause increased vascular permeability, contraction of smooth muscle, and release of histamine from basophils, and mast cells

c1q

• binds to fc of igm and igg

c1r

• activates c1s

c1s

• cleaves c4 and c2

c4

• part of c3 convertase (c4b)

c2

• binds to c4b to form c3 convertase

c3

• key intermediate in all pathway, most concentrated complement component

c5

• initiates membrane attack complex

c6

• binds to c5b in mac

c7

• binds to c5bc6 in mac

c8

• starts pore formation on membrane

c9

• polymerizes to cause cell lysis

factor b

• binds c3b to form c3 convertase

factor d

• cleaves factor b

properdin

• stabilizes c3 convertase and c5 convertase in the presence of magnesium

mbl

• binds to mannose

masp 1

• helps cleave c4 and c2

masp 2

• cleaves c4 and c2

c1 inhibitor

• dissociates c1r and c1s from c1q

factor i

• cleaves c3b and c4b

factor h

• cofactor with i to inactivate c3b, prevents binding of b and c3b

• the principal soluble regulator of the alternative pathway

c4 binding protein

• acts as a cofactor with i to inactivate c4b

s protein (vitronectin)

• prevents attachment to the c5b67 complex to cell membrane

daf (d55)

• dissociates c2b or Bb from binding sites, thus preventing formation of c3 convertase

mirl (cd 59)

• prevents insertion of c9 into cell membrane

cr1 (cd 35)

• cofactor with i; mediates transport of immune complexes

• it is found mainly on peripheral blood cells

• it binds c3b and c4b but has the greatest affinity for c3b

• once bound to cr1, both c4b and c3b can then be degraded by factor i

mcp (cd 46)

• found on virtually all epithelial and endothelial cells except erythrocytes

• is the most efficient cofactor for factor i mediated cleavage of c3b

• it also helps to control the alternative pathway because binding of factor b and c3b is inhibited