Camelid Antibody-Based Therapeutics for Animal and Human Health

Abstract

Modern age has brought about the indiscriminate use of chemicals. Synthetic chemicals are used to increase food production and control diseases with the objective of improving the quality of our lives. Chemicals are not biodegradable and pollute the atmosphere, accumulate inside living bodies, and create a disturbance in the delicate balance of the ecological system governing life on planet earth. This resulted in rampant pollution that is difficult to contain and give rise to drug-resistant microorganisms that are impossible to treat. This has ultimately led to a poor ecosystem and a lowering of the quality of life of all habitants of earth. We humans, who are responsible for this, have woken up to the fact that we need to undo this by developing clean green sustainable technologies and eco-friendly products.

Appendices

Appendix 1: Our Laboratory Activity Chart

Organization Setup Activities

Our laboratory has been started in Jan 2009. Tentative milestones to be attained are shown below:

No.	Item	Date	Comments
1	Preorganization planning activities	Done	Writing up business and scientific plans, Market research and pre business development activities. Determination of collaborators/partners and management and scientific teams
2	Corporate plan review and finalize open issues and negotiations with funding agencies (tie-ups with corporate houses, financial institutions, VCs, Angel Investors, Govt, e.g., DBT, DST, CSIR, and quasi-Govt, e.g., biotech parks, IIT, ILS). Collaborations/partnerships with organizations working in this area if necessary. Entering into JV/agreement with collaborators	In progress	Board of directors and scientific advisory board will be finalized after consultation with all the partners
3	Obtaining the funding and setting up of application laboratory (viz. instrumentation, infrastructure, personnel recruitment)	Oct 2009	Setting up of application laboratory and hiring of personnel with funding from investors. SOPs will be optimized simultaneously
4	Corporate setup completed and begin formal operations and standardization of instruments, testing of HVAC, first meeting to decide the course of work, timelines and deliverables	Dec 2009	Full fledged operational laboratory
5	Full fledged commencement of wet laboratory work (experiments started)	Jan 2010	R&D work commences. Two different project areas, one for probiotics and the other in the area of camelid antibodies begin

Appendix 2: Three-Year Road map

Timelines	Proposed progress chart
Oct–Dec 2009	− Setting up of the application laboratory and making it operational − Hiring personnel and organizing labs to functional domains − Setting up of the probiotic culture facility with a small fermenter − Procuring antigens genes and expression of proteins for immunization of camels/llamas
Jan–April 2009	− Immunization of Llamas − Fine-tuning of protocols for screening of the phage library by panning. Establishing the optimized protocol for the same with a standard library − Organizing blood collection and RNA isolation from camels/llamas − Preparing the camelid library in phage, bacteria, and yeasts with the cDNA from the immunized Llama
May–Aug 2010	− Panning of the library against antigens and selection of the right clones. Transformation in E. coli and selection of the antibody producing clones by ELISA. Calculation of binding affinity − Optimization of the growth conditions for Saccharomyces and Lactobacillus. Establishing the transformation protocol with food grade secretory vectors that would give high yields
Sept–Dec 2010	− Optimization of growth conditions of the probiotic cultures in consortium − Introduction of the first genetically modified yeast in the consortium and accessing if the consortium secreted effective antibodies − Alternatively producing killed yeast and mixing with the probiotic culture to see if they are effective − Finalizing the composition and formulation of the consortium in the value-added probiotic format. Building up of pilot-scale live cultures in various formats and testing for the stability, resistance to contamination, maintenance of the sub populations in the consortium, etc.
Jan–April 2011	− Large-scale manufacture of the product for clinical trials
May–Aug 2011	− Clinical trials, regulatory clearances, and test marketing. Clinical trials will be conducted in various poultry farms in India. These will be the future customers for the product and in a way test marketing will also be done during clinical trials. Clinical trials will be conducted with two types of products: 1. Non-GMO cultures: consortium of probiotics with dead yeast cultures containing the camelid antibodies to toxins 2. GMO cultures: consortium of probiotics containing camelid antibody secreting yeasts Product ready for the market. It can be out licensed or manufactured by our laboratory after a fresh round of funding to set up the manufacturing facility

1. Outline of the green technology development and technology extension