Mouse immunoglobulin

Segal, D.M., et al. The three-dimensional structure of a phosphorylcholine-binding mouse immunoglobulin Fab and the nature of the antigen-binding site. Proc. Natl. Acad. Sci. USA 71 4298-4302, 1974. 

CN immunoglobulin G (human-mouse monoclonal c7E3 clone p7E3VHhCy4 Fab fragment antihuman... 

Bach, M.A, Kohler, H. and Levitt, D. (1983) Binding of phosphorylcholine by non-immunoglobulin molecules on mouse B cells. Journal of Immunology 131, 365-371. 

The most common carrier proteins in use today are keyhole limpet hemocyanin (KLH MW 4.5 X 105 to 1.3 X 107), BSA (MW 67,000), aminoethylated (or cationized) BSA (cBSA), thyroglobulin (MW 660,000), ovalbumin (OVA MW 43,000), and various toxoid proteins, including tetanus toxoid and diphtheria toxoid. Other proteins occasionally used include myoglobin, rabbit serum albumin, immunoglobulin molecules (particularly IgG) from bovine or mouse sera, tuberculin purified protein derivative, and synthetic polypeptides such as poly-L-lysine and poly-L-glutamic acid. 

Procedure for blocking immunoglobulins homologous to primary antibody (e.g., Mouse-on-Mouse )... 

Fig. 9.1 Immunolocalization of Adenosine Receptor A (FITC, green) in mouse myocardium using mouse monoclonal primary antibody after blocking mouse endogenous immunoglobulins by preincubation with unconjugated Fab fragment Goat Antimouse IgG. Red color accounts for cardiomyocytes and erythrocytes autofluorescence captured under illumination with a filter excit ing the autofluorescence in red spectrum. Nuclei are counterstained with DAPI (blue). Courtesy of Stephanie Grote...

Based apparently on the same principle as described above, suitable but rather expensive kits exploiting monovalent Fab fragments to block endogenous tissue immunoglobulins have also been developed commercially, such as Mouse-on-Mouse (M.O.M. ) Kits by Vector Laboratories (http //www.vectorlabs.com/). Three Vector M.O.M. kits are available. These kits use the same blocking technology and biotin-avidin detection format, but offer a choice of using either an enzyme-based or fluorescent-based visualization method. 

A very interesting approach was presented recently by Niemeyer et al. [113]. They prepared covalent DNA-streptavidin conjugates to which biotinylated alkaline phosphatase, beta-galactosidase, and horseradish peroxidase, as well as biotinylated anti-mouse and anti-rabbit immunoglobulins, were coupled. Immobilization of DNA-streptavidin conjugates was performed by hybridization with the complementary oligonucleotides, bound to the surface. It was demonstrated... 

If the product is an antibody, then it is essential to distinguish the immunoglobulin product, e.g., mouse IgG, from any media immunoglobulin components, e.g., bovine IgG. Lucas et al.16 developed an immunoassay to measure nanogram quantities of bovine IgG in the presence of a large excess of a structurally homologous protein, mouse MAb. The bovine IgG was a contaminant that copurified with the product from a protein A column. For the bovine IgG assay, whole IgG and protein A-purified IgG reacted differently in the assay. It is important to evaluate these types of assays for cross-reactivity. For other media components, such as chemicals or antibiotics, ELISA is probably not the most appropriate method due to the low immunogenicity of chemicals. Techniques such as HPLC would be better to detect these chemical components. 

Tanaka S. Takasu Y, Mikura S, Satoh N, Ichikawa A Antigen-independent induction of histamine synthesis by immunoglobulin E in mouse bone marrow-derived mast cells. J Exp Med 2002 196 229-235. 

Magnetic Dynabeads with M280 sheep antibody to mouse immunoglobulin G (IgG Dynal Biotech/Invitrogen). 

---