Methods and vectors for generating antibodies in avian species and uses therefor

Abstract

The present invention relates to processes for producing polyclonal and monoclonal antibodies to an antigen in an avian species, preferably in a chicken, using polynucleotide vaccination. The present invention also relates to processes for determining the proteomics profile of a set of pre-selected DNA sequences isolated from a bio-sample, preferably the proteomics profile of a human cDNA library. The present invention additionally relates to processes for identifying physiologically distinguishable markers associated with a physiologically abnormal bio-sample. The present invention further relates to antibody arrays, integrated databases for identification of genes and proteins, multi-functional gene expression vectors, and methods of producing and using such antibody arrays, integrated databases and multi-functional gene expression vectors.

Description

[0001] This is a continuation-in-part of PCT / US99 / 26843, filed Nov. 12, 1999, now pending, which claims the benefit of priority under 35 U.S.C. § 119(e) to U.S. Provisional Application Ser. No. 60 / 108,487, filed Nov. 16, 1998. The disclosures of the above-referenced priority applications are incorporated by reference in its entirety.TECHNICAL FIELD [0002] The present invention relates to processes for producing polyclonal and monoclonal antibodies to an antigen in an avian species, preferably in a chicken, using polynucleotide vaccination. The present invention also relates to processes for determining the proteomics profile of a set of pre-selected DNA sequences isolated from a bio-sample, preferably the proteomics profile of a human cDNA library. The present invention additionally relates to processes for identifying physiologically distinguishable markers associated with a physiologically abnormal bio-sample. The present invention further relates to antibody arrays, integrated da...

AI Technical Summary

Benefits of technology

[0026] The present invention further encompasses a method for generating an integrated database for identification of genes and proteins, which method comprises: 1) delivering bioimformatic information of the plurality of genomic DNA sequences, the plurality of cDNA sequences, the plurality of dual expression vectors, the plurality of proteins or peptides, and the plurality of avian antibodies obtained using the above-described methods into the corresponding genomic DNA, cDNA, dual expression vector, protein or peptide, and the avian antibody subdatabases; and 2) providing means for connecting the bioimformatic information from one subdatabase to any or all of the other subdatabases. Preferably, the method further comprises a step of delivering bioimformatic information of the immunospecificity of the avian antibodies obtained using the methods described in the following Section C into the integrated database. Also preferably, the method further comprises a step of delivering bioimformatic information of the proteomics profile of the selected and marked plurality of genomic DNA sequences, which can be obtained according to the methods described in the following Section C, into the integrated database.

Problems solved by technology

Third, only small quantities of plasmid DNA are necessary for antigenic stimulation.

This technique is invasive, time consuming and costly, involving restraint of and blood sample collection from the animals.

It can be difficult to raise antibodies of high specificity against well conserved mammalian proteins.

However, within the first 3 weeks post-immunization, no detectable antibodies were found in any of the vaccine groups.

Although the currently available large-format 2-DE is capable of producing gels containing up to 10,000 distinct protein and peptide spots, over 95% of the spots separated by such 2-DE gel cannot be sequenced because they are beyond the limits of current high-sensitivity Edman sequencing technology (Persidis, Nature Biotechnology, 1998, 16:393-394).

However, such vaccination is time consuming and costly.

Therefore, due to the vast number of the known DNA sequences to be characterized, it is virtually impossible to use the conventional protein or peptide vaccination technology to generate antibodies for large-scale proteomic research.

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