Freund s adjuvant

Polyclonal Antibodies against FORL r purified polygalacturonase were raised in white rabbits. For the first immunization 200 jxg of purified protein in 300 jxl of distilled water was mixed with 200 1 of PBS and 500 n of complete Freund s adjuvcmt and injected intramusculary into the leg. Two subsequent intramuscular injections, each containing 300 fig of protein in 1 ml of incomplete Freund s adjuvant were given at 1 month intervals. Finally, the rabbit was bled 1 week later. The antisera, separated from blood by incubation at 37 "C, were stored in 1 ml fractions at -20 C. 

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. 

These results show that the covalently crosslinked Adjuvax formulations were superior to the physically entrapped Adjuvax formulations. In addition, the Adjuvax crosslinked formulations were as effective in stimulating antibody titers as Freund s Adjuvant. Furthermore, animals immunized with Adjuvax did not experience local inflammatory reactions or granuloma formation which was observed with all CFA/IFA immunized animals. 

Figure 4. Comparison of Freund s Adjuvant to Adjuvax formulations in stimulating antibody response to P55 oligopeptide antigen. Relative antibody titers between adjuvant groups at day 27 were determined by measuring the absorbance at 450 nm of a 1 500 dilution of anti-P55 immune sera by ELISA.

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. 

AP immunotherapy has been used in both prevention and treatment trials in mutant mice. Both Ap immunization (with Freund s adjuvant) and passive transfer of Ap antibodies reduce levels of AP and plaque burden in... 

In contrast to certain oil emulsions (including Incomplete Freund s Adjuvant), the liposphere approach uses pharmaceutically acceptable biodegradable constituents. 

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. 

Rabbit Immunization. Six rabbits were sub-cutaneously injected with. 2 mg of the immunogen in Freund s adjuvent on days 0, 12, 26, 40 and 59 of the immunization schedule (see Table I). Complete adjuvent was used for the first three immunizations followed by incomplete adjuvent for the final two immunizations. The rabbits were bled as needed using ear vein puncture. 

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. 

Mirvish, S.S.. Nickols. J.. Weisenburger. D.D., Johnson, D., Joshi, S.S., Kaplan, P, Gross, M. Tong. H.Y (1991) Effects of 2.4.5-trichlorophenoxyacetic acid, pentachlorophenol, methylprednisolone. and Freund s adjuvant on 2-hydroxyethylnitrosourea carcinogenesis in MRC-Wistar rats. J. Toxicol, environ. Health, 32, 59-74... 

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