Antibody products

DB Biotech – 3 Levels of Antibody Quality

DB Biotech can produce an antibody for any existing protein with a known amino acid sequence, disease-related mutation, or post-translational modification. The target protein is analyzed from 3 points of view; the partial primary sequence determines the formation of the efficient target epitope for antibody interaction. Those sequences are further analyzed as a potential portion of interacting domains, regulatory posttranslational modification sites, or as a target for proteolytic cleavage. When this is not the case, the partial peptide sequence is further analyzed from a higher tertiary structure point of view for its accessibility on the protein molecule surface in solutions at different stages of cell signaling pathways.

If a special posttranslational modification is a target, the sequence surrounds the target amino acid(s) with other amino acids respecting the criteria mentioned above. Rabbits are immunized with a synthesized peptide conjugated with the carrier protein.

Three levels (qualities) of antibodies are prepared from crude antiserum:

1) Immunoaffinity purified fraction of specific IgGs suitable in most cases for western blot, immunohistochemistry and ELISAs.
2) In vitro cloning technology produces pure clones of antibodies, each corresponding to a single clone of B lymphocytes. This level of purity has a unique specificity, avidity and affinity. Moreover, the specific activity of this fraction is the highest possibly obtainable. Only a few nanograms of immunoglobulins are needed for one reaction in highly specific quantitative or semiquantitative ELISAs or other antibody-based diagnostic assays.
3) Clonal antibodies can be humanized for human in situ application/drug development. This is basically limited to pathology-related change of target protein molecules, where the pathological mutation and/or posttranslational modification creates a unique and specific new “disease-identifying” epitope.

DB Biotech uses a new company-client approach in antibody design and production. First of all, our company consults the given project and the corresponding customer demands regarding the antibody. The target protein is consequently analyzed from various aspects of complete proteomic analysis. The resulting designated peptide is used for the production of polyclonal peptide antibodies. At the highest quality of preparation, pure clones of immunoglobulins are separated from crude antiserum by newly introduced technology as mentioned above.

The following are several examples of antigens against which clonal antibodies are produced:

• actin
• p53
• iNOS
• γ-Interferon
• Protein Kinase C (PKC)is oforms:
• PKC-α
• PKC-βI
• PKC-βII
• PKC-γ
• PKC-δ
• PKC-ε
• PKC-ζ

Each clonal antibody is analyzed for its affinity and specificity, and a detailed data sheet is provided with all the necessary information regarding its use for western blot, immunochemistry, ELISA and immunoprecipitation.