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PubMed1990

Lipid modification of the 15 kiloDalton major membrane immunogen of Treponema pallidum.

The 15 kiloDalton major membrane immunogen was included among the Treponema pallidum polypeptides selectively labelled with [3H]-palmitate. The cloned gene for this immunogen, tpp15, encoded a signal peptide of 17 amino acids, a consensus signal peptidase II cleavage site, and a mature protein of 124 amino acids (13,967 Daltons). As predicted by the DNA sequence, the recombinant 15 kiloDalton immunogen labelled selectively with [3H]-palmitate, and globomycin inhibited processing of the precursor to the mature polypeptide. While the native and recombinant immunogens are amphiphilic, the 15 kiloDalton immunogen synthesized in a cell-free system was hydrophilic. The covalent attachment of fatty acids appears to be responsible for the amphiphilicity of the immunogen and its membrane attachment.

Acylation↗

PubMed1982

Immunogenicity of four sequential polypeptides in inbred guinea-pigs and their recognition at the T lymphocyte and antibody levels.

Four sequential polypeptides containing equimolar amounts of tyrosine, glutamic acid, alanine and glycine were shown to be T lymphocyte-dependent immunogens in inbred guinea-pigs. Poly (Glu-Ala-Tyr-Gly) was immunogenic only in strain 2 guinea-pigs; poly (Glu-Tyr-Ala-Gly) and poly (Ala-Tyr-Glu-Gly) were immunogenic only in strain 13 guinea-pigs; and poly (Ala-Glu-Tyr-Gly) was immunogenic in both inbred strains. The specificity of immune recognition was probed at the T lymphocyte and humoral levels with the heterologous polypeptides. Only a few cases of heterologous cross-stimulation or cross-reaction were observed, indicating the great selectivity of immune recognition. The results showed considerable variability in immune recognition from animal to animal. Nevertheless, at the T-cell level, cross-stimulation appears to necessitate that the antigen is itself immunogenic in that strain, whereas at the antibody level, cross-reaction is not similarly restricted. The structural basis for mutual recognition of immunogen and antigen at these levels is discussed.

Animals↗

PubMed1977

Experimental scrub typhus immunogens: gamma-irradiated and formalinized rickettsiae.

Scrub typhus immunogens were prepared by exposing infected yolk sac suspensions of Rickettsia tsutsugamushi to various doses of gamma radiation. Mouse lethality was abolished at doses greater than 200 krads, whereas immunogenicity of the suspensions, as shown by mouse protection tests, was diminished relatively little by radiation doses in the 200- to 400-krad range. Using a 300-krad gamma dose to provide a safety factor, immunogens were prepared and their protective capacity was contrasted with formalinized scrub typhus immunogens prepared by conventional techniques. Formalinized suspensions afforded mice only partial protection against intraperitoneal challenge with 1,000 50% mouse lethal doses of the virulent homologous strain and no significant protection against similar challenge with an equally virulent heterologous strain. Using the same strains, radiation-inactivated preparations provided 100% protection against 10,000 50% mouse lethal doses of the homologous strain and 70% protection against challenge with the same doses of a heterologous strain. Neither immunogen was a potent stimulator of antibody production as measured by the complement-fixation test. Cell-transfer studies using inbred mice indicated a role for cell-mediated immunity after vaccination with gamma-irradiated immunogens, but no cell-mediated protection could be demonstrated after vaccination with formalin-inactivated rickettsiae.

Animals↗

PubMed1970

Immunogenic mycobacterial ribosomal and ribonucleic Acid preparations: chemical and physical characteristics.

Five to 20% linear sucrose gradients were used to obtain sedimentation patterns of mycobacterial ribosomes, ribosomal subfractions, and ribonucleic acid (RNA) preparations. Classical 70S ribosomes were obtained when 10(-1)m magnesium chloride was used. These, when dialyzed against 10(-4)m MgCl(2), yielded typical 50S, 30S, and smaller ribosomal subunits. The 30S subunits were the most immunogenic under these conditions. A ribosomal preparation containing subunits which varied from 2.5 to 40S was fractionated by collecting five fractions from a sucrose gradient; based upon the amount of nucleic acid present, the fraction containing the 40S particles was most immunogenic. Physical and chemical evidence suggested that mycobacterial RNA preparations extracted with 65% ethyl alcohol from the ribosomes and diluted in distilled water, were either double-stranded, or mostly double-helical, or had a highly organized secondary structure. This was based on the following observations. (i) Native RNA was resistant to trace amounts of ribonuclease. (ii) The approximate T(m) value in SSC buffer (0.15 m NaCl plus 0.015 m sodium citrate) was greater than 85 C and in 0.1 SSC buffer was 55 C; the RNA diluted in SSC buffer produced a hypochromic effect on cooling at room temperature. (iii) Formaldehyde, in the presence of SSC buffer, decreased the T(m) of the RNA to approximately 55 C, and there was no hypochromic effect on cooling. (iv) Formaldehyde did not increase the wavelength of maximal adsorption of the RNA. (v) The purine/pyrimidine ratio was close to one. (vi) The major peak of the RNA sedimented in the more dense zones of the sucrose gradients. There was a relationship between the sedimentation pattern obtained with the RNA-protein subunits on sucrose gradients and immunogenicity; several examples are given. RNA-protein complexes of approximately 14 to 20S, and occasionally 23S in the major peak, appeared to produce the highest immune response. Smaller RNA-protein complexes such as 6S, which were obtained when the RNA preparation was diluted in certain buffers, were much less immunogenic. This was confirmed by collecting five fractions from sucrose gradients and finding the third fraction (containing RNA-protein complexes approximately 15 to 16S) the most immunogenic. Immunogenic activity was apparently related to the structure of the RNA since it was maximal when the RNA appeared to be either double stranded, double helical, or had a highly organized structure.

Journal Article↗

PubMed1978

Pseudomonas ribosomal vaccines: preparation, properties, and immunogenicity.

The preparation, properties, and immunogenicity of ribosomal vaccines from Pseudomonas aeruginosa are described. These preparations, containing protein and RNA, were tested for immunogenicity by active immunization of mice and subsequent challenge with homologous, live bacteria. The results demonstrated that vaccines prepared from a majority of serotypes used were immunogenic, i.e., afforded 60 to 100% mouse protection against a challenge inoculum containing 8 to 50 50% lethal doses. In some cases vaccine doses as low as 1 microgram of RNA provided 100% mouse protection. Molecular sieve chromatography of a highly immunogenic ribosomal preparation on Sepharose 4B demonstrated the presence of two molecular weight fractions: (i) peak A, an excluded peak (thus having a molecular weight of at least 2 times 10(7)), and (ii) peak B, considerably retarded, with an elution position corresponding to a molecular weight of about 2.2 X 10(6), approximating that of typical 70S ribosomes. Both peaks A and B were immunogenic; however, the immunogenicity of peak A was greater (i.e., a smaller immunizing dose was required) than that of peak B. Peak A was shown to contain components of lipopolysaccharide in addition to protein and RNA (which comprised 80% of the dry weight of peak A). On the other hand, peak B was shown to be free of lipopolysaccharide, and 100% of its dry weight consisted of protein and RNA.

Animals↗

PubMed1979

Gamma-irradiated scrub typhus immunogens: broad-spectrum immunity with combinations of rickettsial strains.

Scrub typhus immunogens were prepared from Rickettsia tsutsugamushi strains Karp, Kato, Gilliam, Kostival, and Buie by exposing frozen infected yolk sac suspensions to 300 krad of gamma radiation. Mouse protection tests showed that each of the irradiated immunogens protected C3H/HeDub mice against high challenge levels of Karp and Gilliam, but that none of these single-strain immunogens were capable of protecting against all five of the challenge strains. Broad-spectrum protection was achieved by using combinations of three strains of irradiated rickettsiae in a vaccination regimen of three injections at 5-day intervals. A comparison of vaccination efficacy employing three such combinations (Karp-Gilliam-Kato, Karp-Kostival-Kato, and Buie-Kostival-Kato) indicated that both sequential administration of strains on successive vaccination days and multiple injections of trivalent mixtures produced protective responses superior to those obtained with single-strain immunogens. Trivalent mixtures of rickettsiae exhibited a striking synergistic effect on the immune response of C3H/HeDub mice and elicited a protective response against Kato challenge that could not be obtained with any single-strain immunogen. Mice vaccinated with the trivalent Karp-Gilliam-Kato immunogen resisted challenge with more than 10(3) 50% mouse lethal doses of Karp and Gilliam for 12 months, and were resistant to similar levels of challenge with Kato and Buie for 6 months.

Animals↗

PubMed1986

Monoclonal antibody to a major surface glycoprotein immunogen differentiates isolates and subpopulations of Trichomonas vaginalis.

To produce monoclonal antibodies (MAbs) to highly immunogenic membrane proteins of Trichomonas vaginalis NYH286, the sera of subcutaneously infected BALB/c mice were first monitored for antibody to trichomonad surface proteins. The sera possessed antibody to one major surface protein by 7 days and antibody to numerous other trichomonad membrane proteins by 4 weeks postinfection. A hybridoma was then generated that synthesized an MAb, designated C20A3, which reacted to a parasite-derived glycoprotein possessing a molecular weight of 267,000 (267K glycoprotein). The immunogen corresponded to the single high-molecular-weight immunogenic surface protein recognized by 7-day mouse antisera. The MAb differentiated T. vaginalis isolates by a whole-cell enzyme-linked immunosorbent assay and by indirect immunofluorescence, using either fixed or live organisms. All isolates, however, possessed C20A3-reactive material when tested by enzyme-linked immunosorbent assay, using detergent extracts of the isolates incubated with MAb-coated microtiter well plates. The epitope was accessible to antibody binding on live T. vaginalis organisms expressing the major immunogen, and the 267K glycoprotein was readily removed from the parasite membranes by trypsinizing the intact trichomonads. The antigen incorporated radiolabeled glucose, mannose, and acetate. Also, an unlabeled 267K glycoprotein on nitrocellulose blots was detected by 125I-concanavalin A and 125I-wheat germ agglutinin, confirming the glycoprotein nature of the immunogen. Finally, of seven isolates used in this study, one possessed a cross-reactive 170K, rather than 267K, antigen. The data reinforce the idea that antigenic heterogeneity among T. vaginalis isolates may be a function of the presence or absence of high-molecular-weight glycoprotein immunogens from trichomonal membranes.

Animals↗

PubMed1972

The primary immune response in mice. 3. Retention of sheep red blood cell immunogens by the spleen and liver.

The antibody-inducing activities of foreign red blood cell immunogens sequestered in the spleens and livers of mice injected with sheep erythrocytes were evaluated during the early periods of the immune response. Estimates of immunogenicity, obtained from the magnitudes of anti-sheep red blood cell hemolysin responses evoked in sensitized recipient mice by subcellular tissue fractions prepared from these phagocytic organs, showed that the liver and spleen differ greatly in their handling of this particulate antigen. The liver, functioning primarily as a scavenger organ, destroys completely the immunogenicity of the heterologous erythrocytes in 12 hr. In contrast, the spleen handles foreign erythrocyte immunogens in at least two different ways: approximately 90% of the initially sequestered activity is rapidly destroyed by the spleen in 6 hr, but the remaining activity, associated chiefly with a tissue fraction possessing the sedimentation properties of "light mitochondria," is retained at a significant level for 3 days, and then progressively decreases to a low level until the 7th day. A correlation of the observed changes in the properties of the immunogenic tissue fraction with known cellular events in the spleen stimulated by antigens indicates that the retention of the degradable erythrocyte immunogen is essential for stimulating and maintaining immune reactions in this antibody-producing organ.

Animals↗

PubMed1972

Effect of polybasic amines on the immunogenicity of mycobacterial ribonucleic acid.

The five polybasic amines, methylated bovine serum albumin (MBSA), protamine sulfate, neomycin sulfate, streptomycin sulfate, and diethylaminoethyl-dextran (DEAE-dextran), were examined to determine their effects on the immunogenic activity of mycobacterial ribonucleic acid (RNA) preparations, and whether they could substitute for Freund incomplete adjuvant (FIA) by protecting the mycobacterial RNA in vivo. These compounds were either emulsified in FIA or not emulsified in FIA. Different results were obtained when these compounds, complexed with mycobacterial RNA, were emulsified in FIA. MBSA, in ratios of mycobacterial RNA-MBSA of 1:0.2 to 1:0.4, had no effect on immunogenic activity. However, when the ratio was increased to 1:1, 1:2, or to 1:4, marked inhibition of the immunogenic activity occurred. Protamine sulfate and neomycin sulfate also inhibited the immunogenic activity of mycobacterial RNA; however, neither streptomycin sulfate nor DEAE-dextran had any effect on immunogenic activity. Without being emulsified in FIA, these five polybasic amines, with the exception of DEAE-dextran, acted only as weak adjuvants for mycobacterial RNA and, therefore, could not be used as substitutes for FIA for the protection of mycobacterial RNA from host nucleases. DEAE-dextran, although not as effective as FIA, afforded some protection for the mycobacterial RNA. DEAE-dextran alone also produced a low degree of nonspecific immunity against tuberculosis. Since MBSA, protamine sulfate, and neomycin sulfate reduce the biological activity of mycobacterial RNA after complexing with it, it is probable that these compounds "mask" the immunogenic sites on the mycobacterial RNA structure.

Amines↗

PubMed2002

Differences in components at delayed-type hypersensitivity reaction sites in mice immunized with either a protective or a nonprotective immunogen of Cryptococcus neoformans.

Cell-mediated immunity is the major protective mechanism against Cryptococcus neoformans. Delayed swelling reactions, i.e., delayed-type hypersensitivity (DTH), in response to an intradermal injection of specific antigen are used as a means of detecting a cell-mediated immune (CMI) response to the antigen. We have found previously that the presence of an anticryptococcal DTH response in mice is not always indicative of protection against a cryptococcal infection. Using one immunogen that induces a protective anticryptococcal CMI response and one that induces a nonprotective response, we have shown that mice immunized with the protective immunogen undergo a classical DTH response characterized by mononuclear cell and neutrophil infiltrates and the presence of gamma interferon and NO. In contrast, immunization with the nonprotective immunogen results in an influx of primarily neutrophils and production of tumor necrosis factor alpha (TNF-alpha) at the DTH reaction site. Even when the anticryptococcal DTH response was augmented by blocking the down-regulator, CTLA-4 (CD152), on T cells in the mice given the nonprotective immunogen, the main leukocyte population infiltrating the DTH reaction site is the neutrophil. Although TNF-alpha is increased at the DTH reaction site in mice immunized with the nonprotective immunogen, it is unlikely that TNF-alpha activates the neutrophils, because the density of TNF receptors on the neutrophils is reduced below control levels. Uncoupling of DTH reactivity and protection has been demonstrated in other infectious-disease models; however, the mechanisms differ from our model. These findings stress the importance of defining the cascade of events occurring in response to various immunogens and establishing the relationships between protection and DTH reactions.

Animals↗

PubMed1969

Isolation and partial characterization of an immunogenic moiety obtained from Salmonella typhimurium.

Ribosomal preparations obtained from Salmonella typhimurium by differential centrifugation and sodium dodecyl sulfate (SDS) treatment of the bacillary lysate were found to be immunogenic in F(1) hybrid (C(3)H/HeJ x DBA/2J) and albino Swiss mice, as determined by progressive host survival. The immunity obtained was independent of the need for adjuvant and dependent on the dosage of immunogen given. Immunizations with the ribosomal preparations induced an immune response comparable to that obtained by vaccination with living organisms and significantly greater than that obtained by immunization with heat-killed salmonellae, purified lipopolysaccharide, or crude and SDS-treated endotoxin preparations. No effect on the immunogenicity of the ribosomal fraction was observed by enzymatic treatment with trypsin, Pronase, deoxyribonuclease, and pancreatic ribonuclease. Linear sucrose density gradient resolution of the preparations showed that the immunogenicity of the ribosomal fraction was not unique to any one of its subcomponents. Ethyl alcohol-precipitated, crude ribonucleic acid preparations obtained from the ribosomal and sucrose density-resolved ribosomal preparations were found to induce an immune response comparable to that obtained by immunization with the entire ribosomal fraction. Dialysis in doubly distilled demineralized water slightly reduced the immunogenicity of the preparation; however, comparable dialysis in 10(-4)m MgCl(2)-phosphate buffer did not. Chemical assays of the preparations found to be immunogenic were performed.

Adjuvants, Immunologic↗

PubMed2006

Cross-subtype T-cell immune responses induced by a human immunodeficiency virus type 1 group m consensus env immunogen.

The genetic diversity among globally circulating human immunodeficiency virus type 1 (HIV-1) strains is a serious challenge for HIV-1 vaccine design. We have generated a synthetic group M consensus env gene (CON6) for induction of cross-subtype immune responses and report here a comparative study of T-cell responses to this and natural strain env immunogens in a murine model. Three different strains of mice were immunized with CON6 as well as subtype A, B, or C env immunogens, using a DNA prime-recombinant vaccinia virus boost strategy. T-cell epitopes were mapped by gamma interferon enzyme-linked immunospot analysis using five overlapping Env peptide sets from heterologous subtype A, B, and C viruses. The CON6-derived vaccine was immunogenic and induced a greater number of T-cell epitope responses than any single wild-type subtype A, B, and C env immunogen and similar T-cell responses to a polyvalent vaccine. The responses were comparable to within-clade responses but significantly more than between-clade responses. The magnitude of the T-cell responses induced by CON6 (measured by individual epitope peptides) was also greater than the magnitude of responses induced by individual wild-type env immunogens. Though the limited major histocompatibility complex repertoire in inbred mice does not necessarily predict responses in nonhuman primates and humans, these results suggest that synthetic centralized env immunogens represent a promising approach for HIV-1 vaccine design that merits further characterization.

AIDS Vaccines↗

PubMed2002

Identification of immunogenic hot spots within plum pox potyvirus capsid protein for efficient antigen presentation.

PEPSCAN analysis has been used to characterize the immunogenic regions of the capsid protein (CP) in virions of plum pox potyvirus (PPV). In addition to the well-known highly immunogenic N- and C-terminal domains of CP, regions within the core domain of the protein have also shown high immunogenicity. Moreover, the N terminus of CP is not homogeneously immunogenic, alternatively showing regions frequently recognized by antibodies and others that are not recognized at all. These results have helped us to design efficient antigen presentation vectors based on PPV. As predicted by PEPSCAN analysis, a small displacement of the insertion site in a previously constructed vector, PPV-gamma, turned the derived chimeras into efficient immunogens. Vectors expressing foreign peptides at different positions within a highly immunogenic region (amino acids 43 to 52) in the N-terminal domain of CP were the most effective at inducing specific antibody responses against the foreign sequence.

Animals↗

PubMed2005

Immunogenicity of recombinant fiber-chimeric adenovirus serotype 35 vector-based vaccines in mice and rhesus monkeys.

Preexisting immunity to adenovirus serotype 5 (Ad5) has been shown to suppress the immunogenicity of recombinant Ad5 (rAd5) vector-based vaccines for human immunodeficiency virus type 1 (HIV-1) in both preclinical studies and clinical trials. A potential solution to this problem is to utilize rAd vectors derived from rare Ad serotypes, such as Ad35. However, rAd35 vectors have appeared less immunogenic than rAd5 vectors in preclinical studies to date. In this study, we explore the hypothesis that the differences in immunogenicity between rAd5 and rAd35 vectors may be due in part to differences between the fiber proteins of these viruses. We constructed capsid chimeric rAd35 vectors containing the Ad5 fiber knob (rAd35k5) and compared the immunogenicities of rAd5, rAd35k5, and rAd35 vectors expressing simian immunodeficiency virus Gag and HIV-1 Env in mice and rhesus monkeys. In vitro studies demonstrated that rAd35k5 vectors utilized the Ad5 receptor CAR rather than the Ad35 receptor CD46. In vivo studies showed that rAd35k5 vectors were more immunogenic than rAd35 vectors in both mice and rhesus monkeys. These data suggest that the Ad5 fiber knob contributes substantially to the immunogenicity of rAd vectors. Moreover, these studies demonstrate that capsid chimeric rAd vectors can be constructed to combine beneficial immunologic and serologic properties of different Ad serotypes.

Adenoviridae↗

PubMed2006

Safety and immunogenicity of a candidate vaccine for visceral leishmaniasis (Alum-precipitated autoclaved Leishmania major + BCG) in children: an extended phase II study.

BACKGROUND: Untreated visceral leishmaniasis (VL) is an inevitably fatal childhood disease. First-generation candidate vaccines for VL [autoclaved Leishmania major (ALM) + BCG] have been found to be safe and immunogenic but not superior to BCG alone. Modulation of ALM by adsorption to Alum significantly increases the immunogenicity. The Alum-adsorbed ALM vaccine was found to be safe and strongly immunogenic in healthy adult volunteers in a non-VL-endemic area. This study aimed at establishing the safety and immunogenicity of Alum-precipitated autoclaved L. major + BCG vaccine in children under field conditions. METHODS: A total of 544 healthy, leishmanin non-reactive children (<15 y) were randomly allocated to receive either a single intradermal injection of Alum/ALM + BCG or vaccine diluent (placebo). Volunteers were closely followed for 2 years at 6-month intervals for vaccine safety and immunogenicity. RESULTS: The vaccine was well tolerated with minimal side-effects. Leishmanin skin test conversion (>or=5 mm) was seen in 56%, 50%, 25% and 31% at 6, 12, 18 and 24 months post-vaccination, respectively; conversion in the placebo group was 4%, 12%, 3% and 13% at the same follow-up visits. There was no significant increase in anti-leishmanial antibodies in either study arm at any of the follow-up visits. During the study, four patients in the placebo arm developed parasitologically confirmed VL. CONCLUSION: Alum/ALM + BCG vaccine is safe and immunogenic in children under field conditions. Multiple injections might be needed to obtain results similar to those obtained in healthy volunteers.

Adolescent↗

PubMed2025

Increased immunogen valency improves the maturation of vaccine-elicited HIV-1 VRC01-like antibodies.

Antibodies belonging to the VRC01-class display broad and potent neutralizing activities and have been isolated from several people living with HIV (PLWH). A member of that class, monoclonal antibody VRC01, was shown to reduce HIV-acquisition in two phase 2b efficacy trials. VRC01-class antibodies are therefore expected to be one component of an effective HIV-1 vaccine elicited response. In contrast to the VRC01-class antibodies that are highly mutated, their unmutated forms do not engage HIV-1 envelope (Env) and do not display neutralizing activities. Hence, specifically modified Env-derived proteins have been designed to engage the unmutated forms of VRC01-class antibodies, and to activate the corresponding naïve B cells. Selected heterologous Env must then be used as boost immunogens to guide the proper maturation of these elicited VRC01-class antibodies. Here we examined whether and how the valency of the prime and boost immunogens influences VRC01-class antibody-maturation. Our findings indicate that, indeed the valency of the immunogen affects the maturation of elicited antibody responses by preferentially selecting VRC01-like antibodies that have accumulated somatic mutations present in broadly neutralizing VRC01-class antibodies isolated from PLWH. As a result, antibodies isolated from animals immunized with the higher valency immunogens display broader Env cross-binding properties and improved neutralizing potentials than those isolated from animals immunized with the lower valency immunogens. Our results are relevant to current and upcoming phase 1 clinical trials that evaluate the ability of novel immunogens aiming to elicit cross-reactive VRC01-class antibody responses.

AIDS Vaccines↗

PubMed2002

Isolation of immunogenic outer membrane proteins from Mannheimia haemolytica serotype 1 by use of selective extraction and immunoaffinity chromatography.

OBJECTIVE: To use antibodies produced by calves in response to infection with Mannheimia haemolytica in immunoaffinity chromatography for the identification and subsequent isolation of the dominant immunogenic antigens from bacteria grown in iron-deficient media. SAMPLE POPULATION: Serum from 10 calves actively infected with M haemolytica. PROCEDURE: An outer membrane protein fraction was obtained from sonicated salt-extracted M haemolytica cells by extraction with N-lauroyl sarcosinate. The immunoglobulin fraction of serum from calves actively infected with M haemolytica was used to prepare an immunoaffinity column. The immunoaffinity column was used to isolate the dominant immunogenic proteins from the outer membrane protein fraction. The resultant immunogenic protein fraction was subjected to ELISA and immunoblot methods as well as carbohydrate quantification. Sequencing of the N-terminal was performed on the most prominent protein. RESULTS: 5 immunogenic proteins with molecular weights of 42, 30, 24, 20, and 15 kd were isolated. The immunogenic protein fraction was found to contain 51% carbohydrate. The immunoaffinity column capacity was 1 microg of immunogenic protein/mL of gel. The N-terminal sequence of the 42-kd protein was Tyr-Gln-Thr-Tyr-Gln-Ser-X-Leu-Gln, where X could not be identified. CONCLUSIONS AND CLINICAL RELEVANCE: lmmunogenic proteins were isolated by use of immunoaffinity chromatography. A substantial amount of carbohydrates was co-purified in the process. Additional experiments are needed to determine whether the carbohydrates would hinder or enhance development of vaccine preparations. This method could potentially allow a more rapid production of antigens for use in vaccines.

Amino Acid Sequence↗

PubMedDate not supplied

Effects of pH and lysine during detoxification of a hemorrhagic principle of Habu snake (Trimeresurus flavoviridis) venom with formalin on the immunogenicity of the toxoid.

A hemorrhagic principle (HR1B) isolated from the venom of Habu snake (Trimeresurus flavoviridis) was detoxified with formalin at pH 5, 7 and 9. Higher pH shortened the period needed for detoxification and lowered the final formalin concentration required for complete detoxification. The extent of polymerization and the immunogenicity of the toxoid formed were dependent upon pH during detoxification; the lower the pH, the higher the extent of polymerization and the immunogenicity of the toxoid. The higher immunogenicity possessed by the toxoid with higher extent of polymerization was confirmed by fractionation of the toxoid prepared at pH 7 on Sephadex G-200 and by comparing the immunogenicity of the toxoid fractions. The presence of lysine during detoxification brought about a partially polymerized toxoid having new immunogenic characteristics and a higher immunogenicity.

Animals↗