Monospecific polyclonal antisera

Carefully selected monoclonal antibodies against apo(a) are well suited for most immunoassays (LI, L2, W17). Precipitation techniques such as nephelome-try or turbidimetry require monospecific polyclonal antisera. 

Monospecific polyclonal antisera against 16 kDa CF II do not crossreact with any other thylakoid polypeptide (Western immuno blot and ELISA [6]). 

Figure 13 Purification of monospecific rabbit antibody against annexin VI. The rabbit polyclonal antibody against rat annexin Iv (CCBP 65/67) reacts against other annexins (CBP 35 and CBP 33). Monospecific polyclonal antibodies against annexin VI were prepared by immobilized antigen affinity chromatography. Purified annexin VI is immobilized on an epoxy activated CIM disk (BIA Separations) and 1 mL of antiserum is diluted 1 5 in PBS and applied to the disk (flow rate 3mL/min). Bound antibodies were eluted by 0.1 M glycine HCI, pH 3.0, in a step gradient. The Western blot analysis of EDTA extracts of Morris hepatoma rat plasma membranes with the antiserum before and after the affinity purification is shown.

The molecular cloning of CVF was accomplished by using a A,gtll cDNA library prepared from a poly-A RNA from freshly obtained cobra venom glands (22). The screening of this expression library was performed with a monospecific polyclonal goat anti-CVF antiserum. 

Monoclonal antibodies can be raised to defined antigens and provide very specific probes (17) practical protocols are provided by Cuello and Cote (18). It is important to keep in mind that monoclonal antibodies— in spite of their monospecificity—do not circumvent the problem of crossreactivity with imi-dentified tissue antigens. The same rigorous testing needs to be performed for monoclonals. To increase the specificity of polyclonal antisera, one can perform affinity purification and/or separate IgGs from the whole serum. For such purifications, commercial columns are available. Details on antiserum/antibody production have been given in many accounts fe.g., 13,18-23). 

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