For Research Use Only Provide high specificity monoclonal antibody in a short period!

Custom Rabbit monoclonal antibody production Service using Ecobody technology

Service Overview

Ecobody technology is a rapid monoclonal antibody screening method, which was invented by a research group headed by Prof. Nakano Hideo, Graduate School of Bioagricultural Sciences, Nagoya University. In this technique, the gene that encodes monoclonal antibody which is isolated from single B-cells was amplified, then synthesized Fab (fragments of antigen binding) antibody by cell-free protein synthesis to acquire target antibody that carries intended specificity. It had been a challenge to isolate monoclonal antibody from B-cells, because of low efficiency and a technical difficulty.
The Ecobody technology overcomes these difficulty, and highly specific monoclonal antibody for the target antigen can be isolated from B-cells that are obtained from small amount of blood in short period of time.


Fig.1 Monoclonal antibody production and its assessment using Ecobody technology

In this custom service, customers will send us whole blood (5 - 10mL) or lymphocyte (106 cells) that are isolated from immunized animals. We will isolate B-cells, then select monoclonal antibody that shows high specificity against the antigen. At the Cosmo Bio in-house laboratory, we can also accept custom service for design and synthesize peptide antigen, and immunization against rabbits.

Table 1 Superiority of Ecobody technology
  iBody
Ecobody technology		 Competitor A
Hybridoma method		 Competitor B
Phage display method
Immunized animal Rabbit Mouse Not used
Period Minimum 2 days* Several months 7 weeks
Culture handling N/A Necessary Necessary (E.coli)
Main technique Antibody expression in the test tubes Cell fusion · Cultivation Expression in the E.coli
Efficiency of antibody acquisition > 70% non-predictable 90%
Acquisition of native antibody Possible Possible Difficult
Provided antibody Format Modified Fab (Possible to swich into IgG) IgG Fab
Quantity Multiple Several kinds Several kinds
Right Disclaim Disclaim Not disclaim

Operation procedure


Fig.2 Work flow for the rabbit mAb production by Ecobody technology.

Step 1. Antigen preparation, Immunization, Blood collection

Turn around time: about 8-12weeks (when "custom peptide synthesis service" by CosmoBio is used.)

  • Antigen preparation, immunization, Blood collectionat the mid-term: Immunize rabbits (NZW or JPW strain).
  • Blood collection and Lymphocyte separation: collect whole blood (5-10mL) into heparin tube. Separate lymphocyte fraction by density gradient centrifugation, transfer it into cell storage solution, and then freeze to store.
  • Ship whole blood (5-10mL) or lymphocyte (106 cells), and fluorescent modified antigen (about 1 mmol: FITC or Alexa647 derivativs) to iBody Inc.

Step 2. FACS selection, Antibody gene amplification, Antibody expression, and ELISA

Turn around time: about 1-2 weeks (iBody Inc.)

  • FACS selection: using magnetic beads and fluorescent modified antibody, select positive cells, then isolate single cells.
  • Amplify antibody gene: amplify the gene by RT-PCR, then confirm by electrophoresis.
  • Antibody expression and ELISA: express amplified antibody genes using cell-free protein synthesis system, then evaluate by ELISA.
    * The number of antigen can be increased as an option.
    * Please consult about the kinds of ELISA system.
  • Provide electrical data to the customer.

Step 3. Select 5 clones

Turn around time: about 1-2 business day (by the customer)

  • We ask to select 5 clones based on the ELISA data.
    * The number of clones can be increased as an option.

Step 4. Sequencing analysis, and delivery of goods

Turn around time: about 1 week (iBody Inc.)

  • Prepare plasmids, and then determine their sequences.
  • Provide sequencing data, and the report.
    * Plasmid will be provided by request.

The samples to provide

Case I. If you already have antigen and immunized animals.

  • Fluorescence modified target antigen (about 1 mmol, FITC or Alexa47 derivatives)
  • Antigens that should not show cross reactivity.
  • Whole blood that was collected from immunized animal (5-10mL) or lymphocyte (106 cells)

Case II. If you have only antigen.

  • Target antigen.
  • Antigens that should not show cross reactivity.

Case III. If you don't have enough antigen.

  • If the antigen is commercially available, please contact us.

Deliverables

  • Antibody activity assessment data, which evaluated by ELISA.
  • Antibody genes' sequencing results for 5 clones.
  • Plasmids for E.coli expression system that encode antibody genes.

* iBody Inc. does not provide antibody that produced during this service to the customer. If you anticipate obtaining antibody, please contact us.

* If this service is used for research purpose, it is prohibited to disclose monoclonal antibody gene and amino acid sequence information in publications or in conference presentation. This is to prevent commercial use by the third party. The memorandum should be singed between the customer and iBody Inc. when order is accepted.

This product is "Research reagent". It is prohibited to use against humans or animals in medical care, clinical diagnoses, or as food.

Back to top