Complete freunds adjuvant

Materials. Microspherical PGG glucan (Adjuvax, Alpha-Beta Technology, Worcester, MA) was prepared from Saccharomyces cereviseae strain R4 cells (11). Zymosan, cytochrome c (cyt c), bovine serum albumin (BSA), yeast alcohol dehydrogenase (ADH), Complete Freunds Adjuvant (CFA) and Incomplete Freunds Adjuvant (IFA) were purchased from Sigma Chemical Co. (St. Louis, MO). 

Both remacemide and its des-glycinyl-metabolite inhibit the inflammatory, mechanical hyperalgesia as well as the edema induced by injection of carrageenan or complete Freunds adjuvant into the rat hind paw (Asghar et al.,... 

This test, performed in rats, is the classical model of inflammatory pain. Intraplantar injection of inflammatory stimuli such as carrageenan, kaolin, or complete Freund adjuvants (CFA) induces paw swelling and increased pain sensitivity. As pain stimulus pressure is applied on the inflammed paw and gradually increased until the animal responds by vocalisation or withdrawal of the paw. Analgesics increase the pressure threshold (Randall and Selitto, Arch. Int. Pharmacodyn. 1957, 111, 409-419). 

Figure 4. Binding of sH-lacto-Fl-tetraitol by rabbit serum (R31) in response to immunization with edestin-etNH-lacto-N-tetraose. Antigen in complete Freunds adjuvant was administered on Days 1, 38, and 58 (arrows). The binding assay is performed as described in the legend to Figure 2.

Raghavendra, V., Tanga, F. Y., and DeLeo, J. A. (2004). Complete Freunds adjuvant-induced peripheral inflammation evokes glial activation and proinflammatory cytokine expression in the... 

The first group underwent a conventional immunization procedure (6) in which one half mg of ECPs were administered subcutaneously in Complete Freunds Adjuvant (CFA) on day one and in Incomplete Freunds Adjuvant (ICFA) on day 7. The rabbits were boosted every 14 days for 160 days and serum collected 7 days after each boost. 

Production of Hybridoma Cell Lines. Eight-week-old female BALB/c mice were injected three times intraperitoneally with 500 xg of human serum albumin (HSA) at 2-week intervals. The first injection was in complete Freund adjuvant (1 1, v/v), and the succeeding two injections were given in incomplete Freund adjuvant (1 1, v/v). The mice were allowed to rest for 30-60 days and injected intravenously with 50 xg of HSA. Four days later, spleen cells from these mice were fused with SP2/0-Agl4 myeloma cells by using polyethylene glycol, as previously described (38). Hy-bridomas of interest were cloned by limiting dilution. 

Three month old male New Zealand rabbits were immunized with irradiated B. jararaca venom. Prime injection was of Img in complete Freund adjuvant. The second was of the same venom amount in incomplete adjuvant and the booster consisted of Img toxoid in saline solution. 

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. 

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. 

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