BioMatNet Item: FAIR-CT95-1039 - Processing Technology for Recovery of Recombinant Antibody Produced in Crop Plants

FAIR-CT95-1039
Processing Technology for Recovery of Recombinant Antibody Produced in Crop Plants

To find similar Items, click on a keyword below:
Biological Conversion : Biotechnology : Crops for Pharmaceuticals/Cosmetics : FAIR Area 2.2 - Bioprocessing : Pharmaceuticals/Cosmetics : Process Engineering : Protein/Amino Acid : Separation/Fractionation

	Type of Project	Shared Cost
	Contract No	FAIR-CT95-1039
	Total Cost	1,448,765 ECU
	EC Contribution	992,175 ECU
	Start Date	01/11/96
	Duration	36 Months

Processing technology for recovery of recombinant antibody produced in crop plants

Objectives

The key objective is to provide generic processing technologies for affinity-based product separation of antibody fragments produced in plants, by exploiting small stable peptides that resemble the natural epitope recognised by the antibody.

Technical Approach

The peptides will be selected on the basis that they are inexpensive to produce in bulk, and designed with the requirements of being linked with large-scale supports to allow quick, reliable recovery and also multiple release of the affinity medium.

Expected Results

I. Direct Results

The process development will be modelled using antibody fragments developed against two antigens e.g. a peptide hormone (FSH or LH) and an enzyme glucose oxidase (GOX). The following measurable objectives should be achieved:
genes of appropriately designed antibody fragments will be transfected into tobacco and potato to provide material on a practical scale for process development
peptide antigen analogues will be defined as binding ligands for the antibody fragments
specific affinity purification media, based on appropriately designed peptide antigen analogues, will be developed with a view to capture and release of plant produced antibody fragments in a stable and multiple reusable system
process control and product QC techniques will be developed to monitor and validate the process
a working pilot process will be evolved for which the performance characteristics will be assessed in terms of product yield, reusability/stability, cost efficiency and ease of further scale up

II. Indirect Results

comparative data on peptide ligand selection using phage display and PEPSCAN analysis; assessing the practicality of both structural and linear peptides as binding ligands
understanding the rules relating the structure and function of phage displayed peptides linked to the selection as practical ligands for affinity chromatography
information towards the optimisation of the expression of functional antibody in plant tissue

Applications

Development of cost-effective technologies for the extraction of recombinant proteins from molecular farming programmes in plants, producing stable functionally active products for health care applications.

Results to Date

A prospective affinity process that lays the foundation for the purification of recombinant antibodies has been established. All the key technologies are in place for laboratory scale-up to a pilot scale activity in Year 3. Peptide ligands are defined and coupled to the conventional solid phase matrix Sepharose. Other matrices are under investigation. Recombinant anti-FSH and anti-GOx antibodies are being produced in Pichia pastoris in laboratory scale fermenters so that pilot scale development of the affinity processes can take place. The scFv genes have been formatted in the appropriate vectors for expression in plants. The immunoassay systems are in place for rigorous QC of the systems.

Peptide Hormone

Phage screening for antibody mimitopes

Contacts

Coordinator

Philip PORTER / Unilever Research Biosciences Division / UNITED KINGDOM

EC Scientific Officer

Ciarán MANGAN / European Commission - DG Research Quality of Life Programme - Unit B1.2 / BELGIUM

Participant