Anti-idiotype Monoclonal Antibody Protocol

Animal

10 Balb/c mice or 6 rats or 2 rabbits

Adjuvant

Complete Freund's Adjuvant (CFA) initially, followed by incomplete Freund's Adjuvant (IFA) or our proprietary immunization adjuvant for all subsequent injections except final boost.

Immunogen

5-10 mg purified antibody from customer. Whole molecule of the antibody will be used for immunization.

Immunization

The immunogen is diluted with sterile saline and combined with the appropriate adjuvant. The antigen and adjuvant are mixed thoroughly to form a stable emulsion, which is injected intraperitoneally, providing enhanced immune response from the sustained presence of the immunogen.

Schedule

Day 1	Bleed 0.2 ml (yields 0.1 ml pre-immune serum) 1^st immunization with antigen in CFA or our proprietary immunization adjuvant
Day 14	2^nd immunization with antigen in IFA or our proprietary immunization adjuvant
Day 21	Test bleed and ELISA
Day 35	3^rd immunization with antigen in IFA or our proprietary immunization adjuvant
Day 42	Test bleed and ELISA
Day 56	4^th immunization with antigen in PBS or saline
Day 60	Cell fusion

Fusion

Fusion Test bleed antisera samples are tested by ELISA with the target antibody and a negative control antibody with the same isotype (Provided by customer). Cell fusions are performed on the mice showing desired immune response to the immunogen.

Expansion and Screening

The fused cells are plated into 96-well culture plates and screened by ELISA with the target antibody and a non-related negative antibody for the specific clones. The desired clones are then expanded into 24-well plates and isotyped.

Subcloning, expansion and cryopreservation

The selected clones are then subcloned by limiting dilution until they are monoclonal and stable hybridomas. Two subclones from each parental clone are expanded into culture flasks, and 4-6 vials of cells for each subclonal cell line are cryopreserved.

Antibody Sequencing

For rabbit anti-idiotype antibody services, the selected clones are performed V-region sequencing.

Delivery

The typical delivery includes two vials of frozen cells or one flask of live cells and 5 ml of culture supernatant per clone.

Note:

We provide free cell line storage service for 6 months from the date of cell freezing.

Anti-idiotype Monoclonal Antibody Protocol

Note:

サポート

会社について