• Fisheries and Aquatic Sciences
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• v.18 no.4
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• pp.379-385
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• 2015

We studied the effects of the proprietary prebiotic Immunogen$^{(R)}$ on the growth, hematology and gut microbiota of common carp fingerlings. A basal diet was formulated using common feed ingredients and supplemented with Immunogen$^{(R)}$ at concentrations of 0, 5, 10, 20 and $40g\;kg^{-1}$, each of which was tested experimentally on replicated groups of fish. The trials ran for 8 weeks. Common carp fingerlings with an initial weight of $4.82{\pm}0.05g$ were randomly distributed among the experimental tanks at a stocking density of 25 fish per tank. The experimental diets were provided thrice per day; on each occasion the fingerlings were given a weight of feed that amounted to 4% of fish biomass. At the end of the experimental period, we determined the growth performance, feed conversion ratio, hematological parameters, body composition and gut micro-flora parameters of the test fish. Inclusion of $5g\;kg^{-1}$Immunogen$^{(R)}$ in the diet significantly improved growth performance and feed utilization in comparison with controls. However, the whole-body composition of the fish was not significantly influenced by prebiotic inclusion. Inclusion of $5g\;kg^{-1}$ Immunogen$^{(R)}$ significantly increased the total bacterial and Lactobacillus counts in fish intestines, but these bacterial parameters were significantly negatively impacted by higher concentrations of the prebiotic. Red blood cells counts were increased by prebiotic dietary supplementation at concentrations of 5 and $10g\;kg^{-1}$ prebiotic. Glucose and cholesterol levels were elevated by administration of Immunogen$^{(R)}$. Thus, dietary supplementation with $5g\;kg^{-1}$ Immunogen$^{(R)}$ improved fingerling common carp growth performance and feed utilization, and beneficially influenced the gut microflora

• Archives of Pharmacal Research
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• v.20 no.1
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• pp.46-52
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• 1997

To obtain more sensitive immunoassay for methamphetamine (MA) determination, the optimum condition of enzyme-linked immunosorbent assay (ELISA) was investigated in regard to immunogens, antibody purification methods and coating tracers. Activated MA, N-(4-aminobutyl)methamphetamine (4-ABMA), was conjugated with bovine serum albumin (BSA) or keyhole limpet hemocyanin (KLH) and used as immunogen. The antibodies were purified by protein G chromatography or various immunoaffinity chromatography-linked MA-protein ligands, such as MA-BSA, MA-KLH or MA-ovalbumin (OVA). Each purified antibody was characterized by means of sensitivity and cross-reactivity using the three MA-protein coating tracers, MA-BSA, MA-KLH and MA-OVA. The best sensitivity of each antibody was acquired with the MA-OVA tracer although the tracer concentration and the antibody titer level at optimum condition were varied. The antibody with high titer level did not always yield good sensitivity. At optimum condition, immunoaffinity chromatography-purified antibodies were better for sensitivity and for specificity than protein G-purified antibodies. The cross-reactivity of the purified antibodies seemed to be affected by immunogen structure and showed somewhat different patterns according to the immunoaffinity ligand utilized. These data show that the antibody purification method as well as choice of coating tracer and immunogen is essential for the sensitivity and specificity of EIA; the optimum condition for assay should be discovered using various methods and combinations.

• Korean Journal of Veterinary Research
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• v.10 no.1
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• pp.5-10
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• 1970

Due to the fact that an inactivated anthrax vaccine may lark its immunogenicity and stability of immunogen a number of spore vaccines were exclusively used worldwide. In these studies a number of important factors were emphasized to achieve the following: selection of non or less allergic strain of anthrax bacillus, capsulation of bacteria. obtaining of non sporulating but vegetative organisms, adequate inactivation of B. anthraccis by means of formalin, adsorption of immunogen to aluminum hydroxide gel. Non or less allergic strains of anthrax bacillus which is inactivated with formalin was selected by a hyperimmunization and shock test in rabbits. Obtaining capsular material and vegetative immunogen, a virulent anthrax organisms were cultivated on sodium bicarbonate medium with of without adding of l-alanine in which B, anthracis grew luxuriantly without forming spores. Inactivation was carried out at $37^{\circ}C$ water bath for 3 days after the bacterial culture was mixed with formalin, in a final concentration of two per cent of formalin. Aluminum hydroxide gel was added to the mixture of anthrax and blackleg bacterin. Vaccines were injected guinea pig via subcutaneous or intramusoular route and challenged after three weeks and the possibilities of protection was tested. Throughout the studies. the above mentioned vaccines possibly protected the vaccinated guinea pigs more than 80 per cent compared to that of the controls. This experimental results strongly suggest that the vaccine may possibly applicable to bovine.

• Archives of Pharmacal Research
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• v.21 no.2
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• pp.135-139
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• 1998

High titer rabbit polyclonal antibodies (pAbs) which show a specificity for saikosaponin a (SSA), have been generated. The immunogen used was a conjugate of SSA linked through its glucose moiety to bovine serum albumin by periodate oxidation method. The antibody titers obtained from two rabbits, innoculated with the immunogen, reached a plateau after the fourth and third booster injection, respectively. The specificity of the pAbs was determined by hapten inhibition assays using several SSA-like structures. SSA competitively inhibited the binding of the rabbit anti-SSA pAbs to SSA-ovalbumin on solid phase, a coated antigen on the well. The antibodies showed high specificity to SSA, exhibiting no significant cross-reactivity with any of SSA analogues tested.

• Korean Journal of Veterinary Research
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• v.26 no.2
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• pp.225-228
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• 1986

Progesterone immunogen has been synthesized and its melting point, Rf-value, UV and IR spectrum have been measured to develope the essential step in antisera production against low molecular weight substance. Mixed anhydride reaction was used to conjugate $11{\alpha}$-hydroxy-progesterone with succinic anhydride. Melting point of one intermediate compound was $156^{\circ}C$, and Rf-value was 0.41 in benzene : acetone : methanol (5 : 5 : 2). Maximum absorbance was measured at 242nm and ${\varepsilon}$ was $1.641{\times}10^4cm^2/mole$. Loss of hydroxy group was observed at 3450nm, and carbonyl group was appeared at 1160nm, 1250nm and 2960nm. These results indicated that the intermediate compound was progesterone hemisuccinate. Maximum absorbance of progesterone bovine-serum albumin(BSA) conjugate was observed at 250nm. Molar ratio of progesterone to BSA was average 15.4 on UV spectrum.

• The Korean Journal of Physiology
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• v.28 no.1
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• pp.91-97
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• 1994

In order to produce a specific bombesin antiserum far very sensitive radioimmunoassay, synthetic $[lys^3]-bombesin$ conjugated to bovine serum albumin was subcutaneously injected into guinea pigs. The conjugation was performed using either carbodiimide or gIutaraldehyde as a coupling agent. The antisera were characterized by analysis of Scatchard and Sips plots. The antiserum LBE 2G/2 raised by repeat injection of the immunogen conjugated with carbodiimide showed the titer of 1 : 188,000, very low cross-reactivity to bombesin-like peptides except bombesin, with high affinity constant $(1.64{\times}10^{11}\;M^{-1})$ and high heterogeneity index (0.91). The antiserum LBG 1G/2 produced by repeat injection of the immunogen conjugated with glutaraldehyde possessed the titer of 1 : 43,000, high cross-reactivity to some bombesin-like peptides, high affinity constant $(1.19{\times}10^{11}\;M^{-1})$ and high heterogeneity index (0.79). These results indicate that the antiserum LBE 2G/2 is specific only to bombesin and that the antiserum LBG IG/2 binds to some bombesin-like peptides such as alytesin, gastrin releasing peptide and neuromedin C. The antiserum LBE 2G/2 is sufficient for the very sensitive radioimmunoassay of bombesin.

• Journal of Food Hygiene and Safety
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• v.8 no.4
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• pp.205-214
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• 1993

The monoclonal antibody to chloramphenicol(CAP) was produced to develop an enzyme-linked immunosorbent assay(ELISA) for residual CAP. An immunogen(CAP-BSA) was prepared by immunogen, antibody titer was measured by indirect ELISA. Spleen cells form the immunized mouse were fused with SP2/OAg14 myeloma cells. Among hybridomas selected in HAT media, 6 clones shown high antibody titer to CAP were subjected to cloning by limit dilution, and all of the monoclonal antibodies(MCA1, 2, 3, 4, 5, 7 and 9) produced by each clone were identified as IgG1 by ELISA isotyping analysis. Competitive ELISA condition was established by using the purified monoclonal antibody MCA1 as primary antibody and CAP-HSA conjugate as coating antigen. Standard curve of CAP(n=28) showed that the lowest detection limit of CAP is 20ng/ml level. The cross-reactivities of the 6 monoclonal antibodies showed that CAP sodium succinate. CAP base, P-nitrophenol, and p-nitrobenzyl alcohol were 89∼178, 0.050∼2.237, 0.056∼0.794 and 0.013∼7.939%, respectively. No cross-reactivities were observed with phenylalanine, tyrosine, glutamine, thiamphenicol, neomycin, streptomycin, gentamicin, sulfamethazine, sulfathiazole, chlortetracycline and p-aminobenzoic acid.

• Journal of Food Hygiene and Safety
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• v.31 no.6
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• pp.445-450
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• 2016

Thermal-stable soluble proteins (TSSP) in livestock products has been recently reported. Therefore, the development of antibodies and immunoassay using a TSSP is useful because the presence of TSSP can be measured on processed food. In this study, the existence of TSSPs in pork fat and meat was confirmed and their antigenicity was investigated. The extracts from pork fat and meat by heating method were analyzed by SDS-PAGE with 5% stacking and 12% separating gels. The protein profiles from the raw pork fat and meat extracts (major band ranged 25 to 100 kDa) without cooking and heating treatments were significantly different compared to those from cooked and heated pork fat and meat extracts (several major bands > 100 kDa and < 30 kDa). This meant that non thermal-stable soluble proteins ranged from 25 to 100 kDa may be denaturated to insoluble proteins by cooking and heating treatments, and TSSPs were in pork fat and meat at kept their properties. The confirmed TSSPs were used as an immunogen to investigate their antigenicity. Eight mice (5 mice for pork fat and 3 mice for pork meat) were separately immunized with the TSSPs of pork fat and meat, and the anti-sera obtained from the immunized mice showed high titer values. Polyclonal antibodies against each target protein showed the specific reaction to pork fat and meat, individually. These indicated that TSSP could be used as an immunogen to produce antibodies such as monoclonal and polyclonal antibodies. In addition, antibodies specific to TSSP from pork fat and meat may be used as a bio-receptor in immunoassays for the identification of fraudulent adulteration with pork fat and meat in livestock products.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.407.1-407.1
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• 2002

To develop immunostrip of DDT (4.4'-dichlorodiphenyl-2.2,2-trichloroethane) and its metabolites, DDT derivatives (DDA-. DDHP-. DDCP-. DDHH-. and DDHHAP-) were conjugated to carrier proteins (OVA and BSA) and three DDT derivatives (DDA. DDHP. DDCP) were conjugated to KLH for the use of coating ligand and immunogen. respectively. To screen the immunoreactivity of antibody to DDT derivatives, the coating ligand was evaluated by a competitive ELISA and DDHP-OVA was selected. (omitted)

• Journal of Food Hygiene and Safety
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• v.9 no.3
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• pp.123-131
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• 1994

An enzyme-linked immunosorbent assay(ELISA) was developed for the detection of residual gentamicin(GM) in edible animal products. The immunogen(GM-KLH conjugate) and coating antigen(GM-BSA conjugate) were prepared by coupling GM sulfate to keyhole limpet hemocyanin(KLH) and bovine serum albumin(BSA) in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, respectively. Polyclonal antibody to GM was produced in rabbits(New Zealand White, female) by using the immunogen and the antibody titer was measured by indirect ELISA. A competitive ELISA was developed using GM-bovine serum albumin conjugate as a coating antigen, GM(as standards or sample), polyclonal antibody to GM, secondary antibody conjugated with horseradish peroxidase as an enzyme, and H2O2 and o-phenylenediamine dihydrochloride as a substrate and a chromophore, respectively. The detection limit of GM was 10 ng/ml and the standard curve of GM(n=26) was linear up to 10 $\mu\textrm{g}$/ml in this competitive ELISA system. There were no cross-reactivities of the partially purified antibody between GM and the various antibiotice such as amikacin, benzyl-penicillin, chloramphenicol, erythromycin, furazlidone, kanamycin, neomycin, oleandomycin, streptomycin, sulfathiazole and thiamphenicol(CR50<0.05%)