Custom Polyclonal Antibody Production from Primm Biotech, Inc.

Description

Custom Recombinant Protein & polyclonal antibody

Provide us with the gene sequence/ID

Starting from the nucleotide gene sequence or from a gene ID name/number related to public genomic data-base, PrimmBiotech will provide: isolation of the cDNA, gene cloning, expression of the full length gene or of a selected domain (100-300 aa), production of the recombinant protein as His-Tag fusion in E.coli cells and protein purification. The recombinant protein is then used in our standard immunization protocol in two rabbits. Titer is guaranteed and is verified by Elisa testing against the recombinant protein.

Custom polyclonal antibodies against synthetic peptides

The use of synthetic peptides as immunogens is generally applied where either the complete protein is not available in sufficient quantities to carry out an adequate immunization protocol, or to obtain antibodies able to recognize only specific regions of a polypeptide chain. The selection of the amino acid sequence of the peptide to be used as an immunogen is a crucial step toward the success of such a strategy. In fact, one can not predict, a priori, that antibodies raised against specific peptide sequences are then able to efficiently recognize the intact protein from which the peptide sequence was originally selected. Nevertheless, it is possible to exploit certain parameters based on hydrophilicity, accessibility, and flexibility criteria, together with considerations of T-cell epitopes to judge the possible immunogenic power of a given peptide, and thus to increase the possibility that anti-peptide antibodies do recognize the complete and often native conformation of the protein chain. Our technical staff will assist our customers with the selection of the best candidate peptide sequences to be used as immunogens. Specific sequences will be proposed to our customers and submitted for their approval prior to the start of the chemical synthesis. In addition, in some cases the use of more than one peptide from a given protein chain is recommended. After synthesis the peptides are combined and used as a single antigen toward the preparation of antibodies able to recognize all the peptides. Through such an approach, the resulting antibodies have a higher capability to recognize the intact and native protein chain. Generally, synthetic peptides used as immunogens are synthesized on a small scale (5 - 10 mg) and are made of approximately 15 amino acid residues. Small molecules, such as peptides, have low immunogenic potential. To increase their immunogenicity, it is necessary to link, by chemical conjugation, the peptide sequence to a larger protein molecule, which serves as a carrier. Several carrier proteins can be selected to prepare the desired peptide conjugate and, in general, the few required features of the carrier are its molecular weight, which should be higher than 20,000 daltons, and its immunogenic potential. Most of the immunopeptides produced by Primm are conjugated with carriers such as OVALBUMIN or KLH. Normally we conjugate about 3/5 mg of peptide.