• Journal of Plant Biology
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• v.36 no.1
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• pp.91-96
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• 1993

Phycoerythrins from the ceramiaceous red algae Campylaephora crassa (Okamura) Nakamura and related species, C. hypnaeoides J. Agardh and Ceramium kondoi Yendo, were investigated for absorption spectra, protein bands by gel electrophoresis and antigenic reactivity to anti-phycoerythrin using Ouchterlony double diffusion and immunoblot. Similarities in absorption spectra, showing peaks at ca. 566 nm>534 nm>495 nm, were found between C. crassa and Cm. kondoi, while C. hypnaeoides differed slightly. There were no differences in fluorescence emission spectra and protein bands between C. crassa and related species tested. Since Ouchterlony double diffusion, however, showed that phycoerythrins from C. crassa and Cm. kondoi were similar in antigenic reactions, and differed from that of C. hypnaeoides, the taxonomic position of C. crassa should be reinvestigated using other experimental approaches.

• Parasites, Hosts and Diseases
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• v.40 no.2
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• pp.83-88
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• 2002

The 8 kDa antigenic protein of Clonorchis sinensis was partially purified by ammonium sulfate precipitation and subsequently by a column chromatographic steps. The purified protein was separated into 7 and 8 kDa protein bands through SDS-tricine gel electrophoresis, while the protein was fecund to migrate to a 8 kDa band in 7.5-15% SDS-PAGE. The molecular weight of the antigen was estimated to be 110 kDa by Superose 6 HR 10/30 gel filtration. The purified antigen strongly reacted with the human sera of clonorchiasis. The hyperimmune sera of BALB/c mice immunized against the 8 kDa protein were reacted with both the crude extract and the excretory-secretory product of adult worms, but not with the metacercarial extract. Immunohistochemical staining demonstrated that the protein was distributed to the tegument and subtegumental cells and also to the seminal receptacle. The present findings suggest that the 8 kDa protein is a partition of the multicomplek protein originating from various organs of adult C. sinenis, and that it is composed of several 7 and 8 kDa proteins.

• Parasites, Hosts and Diseases
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• v.26 no.3
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• pp.163-168
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• 1988

To observe the antigenic protein fractions in saline extract of Spirometra mansoni plerocercoid (sparganum), the crude extract was separated in reducing conditions of sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE). The proteins, transferred by celctrophoresis to introcillulose paper, were reacted with sera from 15 surgically confirmed sparganosis and 24 cysticercosis patients for immunoblotting. Out of 30 identified protein bands in the extract, bands of 29 and 36 kilodaltons (kDa) were the strongest and the most frequently reacting with specific antibody (IgG) in sparganosis sera. Bands of highter molecular weight also reacted with the sera but their frequency of reactions was lower. Sera of cysticercosis reacted with different protein bands in saline extract of sparganum, but the cross reactions were observed in strong antigenic bands of 29 and 36 kDa.

• Applied Microscopy
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• v.27 no.4
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• pp.403-416
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• 1997

In order to observe the localization of the specific antigenic protein in the tissue of Paragonimus westermani metacercaria, immunogoldlabeling method was applied using IgG of the dog which were infected with Paragonimus westermani metacercaria and IgG of rabbits which were immunized with purified 23 kDa protein from metacercaria of the Paragenimus westermani. The metacercaria worm tissues obtained from Cambaroides similis were embedded in Lowicryl HM20 medium, treated with infected and immunized IgG and protein A gold complex (particle size; 12 nm) and observed by electron microscope. In the tissue antigen of Paragonimus westermani metacercaria, the content of excretory bladder which was highly dense electron density was constituted in the excretory bladder of the parenchymal tissue. In the metacercaria tissues antigen reacted with IgG of infected dog. Labeled gold particles distributed on the interstitial matrix of parenchymal cells, fibrous granules of parenchymal tissue and the content of excretory bladder. High antigenicity was observed on content of excretory bladder. It was found to be specifically distributed at the tissue of Paragonimus westermani metacercaria. In the tissues antigen reacted with IgG of immunized rabbit. Labeled gold particles randomly distributed on the interstitial matrix and fibrous granules of parenchymal tissue but in the content of excretory bladder of Paragonimus westermani metacercaria, gold particles were richly labeled. Therefore, the 23 kDa protein contained with Paragonimus westermani metacercaria was found protein which was specifically constituted at the content of excretory bladder of Paragonimum westermani metacercaria. The 23 kDa protein was commonly contained from of Paragonimus westermani metacercaria to adult and showed strong antigenicity against the immunized and infected IgG.

• The Journal of Korean Society of Virology
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• v.28 no.2
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• pp.139-146
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• 1998

Recombinant polioviruses have been developed by many research groups for use as vaccine vector because poliovirus induces mucosal immunity as well as humoral immunity through oral uptake. We assessed the potential use of poliovirus as a T-cell epitope carrier. Recombinant poliovirus V129 5L was constructed to have a substituted T-helper epitope from the core protein of Hepatitis B virus at neutralization antigenic site 1 on its VP1 capsid protein. The recombinant virus replicated less efficiently than type 1 poliovirus Mahoney strain. The V129 5L formed a little smaller plaques than the Mahoney strain and showed some 1.25 log unit lower titer at the peak in the one-step growth kinetics though it had similar growth profile to that of the Mahoney strain. Since V129 5L recombinant virus was genetically stable even after 24 successive passages in HeLa cells, the antigenic site 1 on VP1 capsid protein was confirmed for its ability of carrying T cell epitope. The genetic stability of V129 5L also indicated that recombinant poliovirus can be successfully utilized for the development of the multivalent vaccines.

• Journal of fish pathology
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• v.22 no.2
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• pp.173-177
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• 2009

Lymphocystis disease virus (LCDV) was detected from olive flounder Paralichthys olivaceus, painted glass fish Chanda baculis, gourami Trichogaster leeri and rockfish Sebastes schlegeli, and proteins of the viruses were compared. The major capsid protein (MCP) gene-specific primer sets successfully amplified approximately 1300 bp nucleotides from the olive flounder and 600 bp nucleotides from painted glass fish, gourami and rockfish isolates, respectively. In western blotting analysis using anti-LCDV mouse polyclonal serum, major antigenic proteins had 21, 26, 45, 50, 80, 110 and 120 kDa in olive flounder, 26, 47 and 80 kDa in painted glass fish, 26, 46, 80 and 92 kDa in gourami, 26, 44, 49, 80 and 105 in rockfish, respectively. All the marine and freshwater isolates showed only common antigens of approximately 26 kDa and 80 kDa. These results suggest that antigenic protein profiles of LCDVs may vary depending upon fish species.

• Pediatric Infection and Vaccine
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• v.6 no.2
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• pp.219-233
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• 1999

Purpose : Respiratory syncytial virus(RSV) is the major cause of lower respiratory tract infection in infants and young children. This study was performed to analyze antigenic and genetic variation of G protein of subgroup A RSV. Methods : One hundred seventy-nine strains isolated at the Seoul National University Children's Hospital over 8 years-period from 1990 through 1998 were analysed for antigenic and genetic variability. Analysis was made by reactivity with monoclonal antibodies raised against RSV, and by restriction mapping and, for selected strains, nucleotide sequencing following amplification of full sequence of G gene by reverse transcription-polymerase chain reaction. Results : Restriction fragment analysis of the amplified G protein gene revealed 23 restriction patterns, 12 of which included more than 2 isolate, and the most frequent genetic type comprised 30% of the strains. Indirect immunofluorescent staining with monoclonal antibodies revealed 6 antigenic types with one predominant pattern accounting for 91% of the total strains. The most frequent antigenic type had 21 restriction patterns, and some viruses with same restiction pattern had different monoclonal antibody reaction pattern. Nucleotide sequence homology of subgroup A was 91~93% between reference(A2, Long) and Korean isolates, 93~99% among Korean isolates. Maximum-parsimony analysis demonstrated that Korean isolates were distinct from reference strains and subgroup A strains were clustered in 4 groups. Conclusion : The restriction analysis pattern of G protein gene identified greater diversity within subgroup A than was seen with the monoclonal analysis and a variety of antigenic and genetic types of RSV are circulating in Korea which are different from reference strains or strains isolated from other countries.

• Parasites, Hosts and Diseases
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• v.33 no.2
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• pp.101-106
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• 1995

Soluble protein of purified Pneumocvstis cnrinii was prepared from experimentally infected rats. SDS-PAGE of the crude antigen resolved about 20 protein bands from 20 to 200 kDa. Out of thenl, 116 kDa band strongly reacted and 45-55 and 100 kDa bands reacted weakly to the positive reference human serum from U.S.A. Western blot analysis with sera of 130 normal children and 15 newborns in Korea revealed specific IgG antibody reaction to 40-55 and 116 kDa protein bands. Forty percent (40.0%) of the 145 sera were positive with any of the antigenic protein bands of R corinii. The positive rate was 56% in 50 males and 33.3% in 48 females. The protein bands 40-55 and 116 kDa from rat P. carinii were confirmed to cross-react with human sera in Korea.

• Journal of Veterinary Science
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• v.23 no.4
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• pp.55.1-55.12
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• 2022

Background: ASF was first reported in Kenya in 1910 in 1921. In China, ASF spread to 31 provinces including Henan and Jiangsu within six months after it was first reported on August 3, 2018. The epidemic almost affected the whole China, causing direct economic losses of tens of billions of yuan. Cause great loss to our pig industry. As ELISA is cheap and easy to operate, OIE regards it as the preferred serological method for ASF detection. P54 protein has good antigenicity and is an ideal antigen for detection. Objective: To identify a conservative site in the African swine fever virus (ASFV) p54 protein and perform a Cloth-enzyme-linked immunosorbent assay (ELISA) for detecting the ASFV antibody in order to reduce risks posed by using the live virus in diagnostic assays. Method: We used bioinformatics methods to predict the antigen epitope of the ASFV p54 protein in combination with the antigenic index and artificially synthesized the predicted antigen epitope peptides. Using ASFV-positive serum and specific monoclonal antibodies (mAbs), we performed indirect ELISA and blocking ELISA to verify the immunological properties of the predicted epitope polypeptide. Results: The results of our prediction revealed that the possible antigen epitope regions were A23-29, A36-45, A72-94, A114-120, A124-130, and A137-150. The indirect ELISA showed that the peptides A23-29, A36-45, A72-94, A114-120, and A137-150 have good antigenicity. Moreover, the A36-45 polypeptide can react specifically with the mAb secreted by hybridoma cells, and its binding site contains a minimum number of essential amino acids in the sequence 37DIQFINPY44. Conclusions: Our study confirmed a conservative antigenic site in the ASFV p54 protein and its amino acid sequence. A competitive ELISA method for detecting ASFV antibodies was established based on recombinant p54 and matching mAb. Moreover, testing the protein sequence alignment verified that the method can theoretically detect antibodies produced by pigs affected by nearly all ASFVs worldwide.

• Journal of Plant Biotechnology
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• v.32 no.4
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• pp.233-241
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• 2005

Plants have considerable advantages for the production of antigenic proteins because they provide an inexpensive source of protein and an easy administration of vaccine. Since a publication describing edible plant vaccine of HBsAg in 1992, a number of laboratories around the world have studied the use of plants as the bioreactor to produce antigenic proteins of human or animal pathogens. Over the last ten years, these works have been mainly focused on three major strategies for the production of antigenic proteins in plants: stable genetic transformation of either the nuclear or plastid genome, or transient expression in plants using viral vectors. As many antigenic proteins have been expressed in tobacco, also several laboratories have succeeded to express genes encoding antigenic proteins in other crop plants: potato, tomato, maize, carrot, soybean and spinach. At present many works for the production of edible plant vaccine against bacteria-mediated diseases have mostly performed the studies of enterotoxins and adhesion proteins. Also the development of new-type antigens (pili, flagella, surface protein, other enterotoxin and exotoxin etc.) is required for various targets and more efficacy to immunize against microorganism pathogens. Many works mostly studied in experimental animals had good results, and phase I clinical trial of LTB clearly indicated its immunogenic ability. On the other hand, edible plant vaccines have still problems remained to be solved. In addition to the accumulation of sufficient antigen in plants, human health, environment and agriculture regulation should be proven. Also oral tolerance, the physiological response to food antigens and commensal flora is the induction of a state of specific immunological unresponsiveness, needs to be addressed before plant-derived vaccine becomes a therapeutic option.