Freund’s adjuvant, complete

Compared to wild-type mice, in TRPVl gene-deleted (—/—) mice, complete Freund s adjuvant evokes significantly less oedema, hyperalgesia and arthritis score [149]. 

AIA, adjuvant-induced arthritis CFA, complete Freund s adjuvant CIA, collagen-induced arthritis DTH, delayed-type hypersensitivity LPS, lipopolysaccharide. 

Fig. 15 Induction of cellular immunity by subcutaneous immunization with OVA-encapsulating y-PGA-Phe nanoparticles. Mice were subcutaneously immunized one time with OVA alone (10 pg), 10 pg of OVA and 100 pg of NPs (OVA-NPs), 10 pg of OVA and 100 pL of complete Freund s adjuvant (OVA + CFA), or PBS (control). Splenocytes were obtained from the immunized mice on day 10 after the immunization and stimulated with the OVA peptide. The number of IFN-y-producing cells was measured by enzyme-linked immunospot assay. SFU spot forming units...

A corollary to the use of cBSA as a carrier protein is that its increased immune response often abrogates the use of complete Freund s adjuvant, which is a source of concern because of its potential side-effects in animals. A relatively innocuous mixture with alum is usually all that is required as adjuvant to result in good antibody production. 

FIGURE 7.7 (See color insert) Adoptively transferred D011.10 transgenicT cells can be identified by expression of CD4+ and KJ-126 in spleen cell suspension from Balb/c mice after ovalbumin (OVA) immunization. Balb/c mice were injected iv with D011.10 spleen cells containing 3-5 x 1 06CD4+KJ-126+ cells and immunized by intraperitoneal injection of 2 mg OVA emulsified in complete Freund s adjuvant 2 days later. OVA immunization increases the frequency of KJ+T cells and alters the expression of various surface molecules consistent with T cell (Tc) activation. 

Antiserum Production The immunogen, carboxymethylmorphine-bovine-serum-albumin, is emulsified with equal volume of complete Freund s adjuvant. Initial immunization doses are injected into the New Zealand albino rabbits and later on this followed up with booster injections after a period of 6 weeks. The antiserum titer is determined with each booster dose injection and is duly harvested when the titre value is maximum. This is diluted suitably and employed in the radioimmunoassay. ... 

Antibody Production A thick emulsion of the immunogen (clonazepam-bovine-serum-albumin-con-jugate) is prepared employing complete Freund s adjuvant and two New Zealand white female rabbits are immunized intradermally at multiple sites with the immunogen emulsion. The animals are then administered... 

Immunization and Antibody Production The immunogen 3-hemisuccinyloxyflurazepam, is emulsified with complete Freund s adjuvant. It is injected intradermally into two female New Zealand albino (white) rabbits. Repeated doses are administered twice at interval of two weeks. Subsequently, booster injections of the thick-immunogen-emulsion-paste are administered after a span of 6-weeks. The antibody is harvested when its titer level is high enough, diluted to the suitable-level and employed in the RIA. 

Immunization and Antibody Production The lypphilized immunogen obtained above is dissolved in normal saline and emulsified with equal volumes of complete Freund s adjuvant into a thick paste. Three New Zealand albino rabbits are immunized with the immunogen-paste through intradermal injections. The process is repeated twice at 2-weeks intervals followed by booster doses at monthly intervals. The antiserum is harvested when the plasma titer value is attained maximum. 

Preparation of Antiserum The barbiturate-bovine-serum-albumin conjugate is duly emulsified with an equal volume of complete Freund s adjuvant and New Zealand albino rabbits are subsequently im-... 

Preparation of Rabbit Serum and Antiserum. Four white New Zealand rabbits (2-3 kg) were first bled to obtain normal serum (NS), then injected subcutaneously with a total of 2.5 mg of either f-1, f-3, f-4, or f-5 in complete Freund s adjuvant. 

Serum antibodies were obtained by immunization with free peptide in complete Freund s adjuvant. 

We have studied the effect of several adjuvants on the antibody responses to several of the peptides (52,53). In general, the highest responses were obtained in immunization of peptide with complete Freund s adjuvant. Immunizations without adjuvant (in PBS) or with Incomplete Freund s adjuvant usually gave little or no responses. 

The first relevance on pain was activation of PPARa in the spinal cord of rats with peripheral inflammation (Benani et al., 2004). Electrophoretic mobility-shift assay was employed to compare the DNA-binding activity toward a PPRE. The PPARa isoform was observed to be activated in the rat spinal cord after complete Freund s adjuvant injection, which could elicit hyperalgesia. PPARa was provided as a new player in the molecular modeling of pain pathways, although it was discussed that inhibitors of PPARa activation might be relevant antinociceptive drugs. 

Various animal species can be used for antibody preparation rabbits are used most often, and goats for larger scale operations. For the first injection using complete Freund s adjuvant, intramuscular injection is recommended, whereas subsequent injections with incomplete Freund s adjuvant are carried out subcutaneously. 

The most popular and successful adjuvants have been the water in oil emulsions developed by Freund. The basic ingredients of light mineral oil (Bayol) and emulsifying agents mixtures such as Arlacel (A or C) are available commercially. The reagents are emulsified with either solutions or suspensions of the immunogen (incomplete Freund s adjuvant). The addition of mycobacteria (Mycobacterium butyricum, M. tuberculosis) in small amounts to the suspension (complete Freund s adjuvant) leads to a further enhancement of the immune response. This has been attributed to the increased local inflammatory response caused by the mycobacteria. ... 

---