Polyclonal & monoclonal custom antibody production services

Custom antibody production - Polyclonal Antibodies

Rockland Immunochemicals has extensive experience in generating high-titer, high-affinity antisera to a wide variety of antigens. We can assist you in experimental design, production, purification and conjugation.

Antigen requirement: Although as little as 200 micrograms of antigen can be used for a standard immunization protocol, we recommend the following:

• If antigen is sparse and valuable, we require 50 micrograms per injection,
• If antigen is abundant, 0.1 to 0.5 mg per injection.

Immunization regime:

No. of animals: 2 or more animals to allow for immune response variability.

Type of antigen: We can immunize animals with native proteins, GST-fusion proteins, peptides, carbohydrates or any other compound of viral, fungal or microbial origin. However, no biohazardous material can be accepted. We recommend the use of Freund's Adjuvant, however, we can accommodate any requirements that you may have for specialized adjuvants.

Conjugation to carriers: We can conjugate carriers like KLH and BSA to enhance the immunogenicity of the peptide.

Supply of antigen: Antigen can be supplied as a lyophilizate, a gel slice or on a nitrocellulose membrane. An additional charge of Rs. 1500/- will be applied for gel slice extraction.

Host selection: We offer polyclonal antibody production in a wide variety of hosts, including Rabbit, Goat, Sheep, Guinea Pig, Chicken, Rat, Mouse, and Donkey.

Custom polyclonal production package:
This service is designed for the rapid screening of peptides usually delivered in about one (1) month! and is the fastest way to get custom polyclonal antibody for your immunoassay.
Includes:

• analysis of the peptide's primary structure
• peptide synthesis (up to 15 amino acids)
• conjugation to KLH carrier protein
• pre-bleeds of two (2) rabbits
• immunization, booster injections
• terminal bleeding of rabbits and titer analysis of the antiserum collected by indirect ELISA
• We will provide you with approximately 100ml of antiserum, the pre-immune serum and any remaining unconjugated peptide.

Monoclonal Antibodies

Through our partners Rockland Inc., USA we offer extensive experience producing monoclonal antibodies and services that are highly customized to your specific requirements. We provide a comprehensive service which includes all steps of monoclonal antibody production from peptide synthesis to ascites production.

Below is a general procedure for production of Monoclonal Antibodies. As each project is unique and experimental needs differ, we at Rockland have the knowledge and experience to help you design the most scientifically sound approach to achieve your goals.

HYBRIDOMA DEVELOPMENT
Service: Phase 1 (Steps 1-3)
Code: MAB-001
Price: Inquire

Step 1: Immunization of 5 Balb/c mice* - In general we begin with a 50 microgram priming dose followed by several boosting doses of 50 microgram each. This regime will continue until a sufficient antiserum titer is achieved.

Step 2: Screening - Test bleeds are routinely performed and evaluated using a direct binding ELISA. Optimal ELISA conditions are developed to ensure the proper evaluation of future clones. In addition test bleeds can be sent to your lab for evaluation in other assays.

Step 3: Evaluation - Data is evaluated to determine the performance of antiserum at this point. Antiserum titers and specific assay performance must be acceptable to continue the project.

Service: Phase 2 (Steps 4-5)
Code: MAB-002
Price: Inquire

Step 4: Fusion - A splenocyte fusion is performed on the mice (2) which responsed the best to the immunizations as determined by ELISA or another assay. The lymphocytes are then fused to an Sp2/0-Ag14 cell line using an optimized PEG mediated fusion protocol.

Step 5: Expansion & Screening - The fusion is plated into a stack of ten to twenty 96 well plates. Plates are monitored for growth and fed weekly. Wells with cell growth are screened by ELISA in 2 to 3 weeks.

Service: Phase 3 (Steps 6-7)
Code: MAB-003
Price: Inquire

Step 6: Subcloning-Cells from five parental positive wells are harvested and subcloned by limiting dilution.

Step 7: Screening & Expansion-Subclones are screened by ELISA, positives expanded, and screened again. Five sublcones will be expanded, frozen and stored for up to six months.

Total Cost for Phase I through III (Steps 1 through 7), please inquire.

Time: Approximately 4-6 months depending on the response to the Antigen.

Antibody Purification

You could purify the following: IgG fraction, IgM fraction, IgY fraction, by Protein A columns, by Protein G columns, by other Affinity Purification methods, HPLC Purification, and Gel Slice Extraction.

Conjugation of antibody

Depending upon your detection requirements, you may wish to conjugate your antibody to the following:
Fluorescein, Rhodamine, Texas Red, Cyanine Dyes, Biotin, Phycoerythrin, Peroxidase, Alkaline Phosphatase, B-Galactoside, Digoxygenin, Peptides, or Biomagnetic particles.

Characterization of antibody

You may wish us to characterize your antibodies by ELISA or Western Blot.