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Biomedical subjects

A A Ilyichev

Publications and source records attributed to A A Ilyichev.

11 recordsLinked to original sources
PubMed2001

Phage antibodies from combinatorial library neutralize vaccinia virus.

The library of human scFv antibodies displayed on the surface of bacteriophages was panned against Vaccinia virus (VACV), strain Elstree. 75% binding with Vaccinia virus. 5 clones were characterized for their binding with VACV and their ability to neutralize VACV in plaque reduction neutralization test (PRNT). Antibodies from the clones were obtained as soluble individual molecules and their binding activities were confirmed in ELISA.

Antibodies, Viral↗
PubMed2000

Insertion of foreign epitopes in HBcAg: how to make the chimeric particle assemble.

Hepatitis B core antigen is one of the most promising protein carriers of foreign epitopes of various human and animal pathogens. Chimeric HBcAg particles can be used as effective artificial immunogenes. Unfortunately, not all chimeric proteins are able to be particulated. The dependence of correct or incorrect folding of chimeric proteins on physical and chemical properties of inserts was studied with the help of ProAnalyst, SALIX and QSARPro computer programs. We have found that insertion of amino acids with high hydrophobicity, large volume, and high beta-strand index prevent self-assembling chimeric proteins. These factors are most important for the C-termini of inserts. Recommendations for obtaining correct folding of chimeric HBcAg particles have been given.

Amino Acid Sequence↗
PubMed1999

Detection of spring viremia of carp virus isolates by hybridization with non-radioactive probes and amplification by polymerase chain reaction.

For detection of spring viremia of carp virus (SVCV) DNA probes have been constructed using the reverse transcription-polymerase chain reaction (RT-PCR) amplification technique and cDNA cloning in plasmid and phage vectors. The specific primers for amplification of SVCV M and G genes were chosen and synthesized. Studies were carried out to establish the sensitivity and specificity of viral RNA detection in infected cell culture and pathogenic material from fish by the use of non-radioactive probes and RT-PCR. The efficiency of amplification with primers, complementary to the genome of the reference Fijan strain, was estimated in RT-PCR experiments with two SVCV strains. Under the same conditions, the quantity of PCR products amplified from the M2 strain was less than that from the ZL4 strain, which implies that the latter is more similar to the reference European SVCV Fijan isolate. Using DNA probes and dot-blot hybridization, SVCV was tested in samples taken from different organs of artificially infected carp with clinical signs of acute disease. The virus could be detected most reliably in fish brain. In most cases the hybridization signal was positive with samples having a viral titer of not less than 10(5) TCID50/g.

Animals↗
PubMed1997

Localization of the amino terminus of the hepatitis B virus core antigen within the core particle.

The position of the amino terminus of the hepatitis B virus core antigen (HBcAg) within the core particle was studied. For this purpose, three recombinant analogs of HBcAg were designed. One analog, HBcAgR, was identical in amino acid sequences to the core polypeptide of the hepatitis B virus; the second, HBeAgR, differed from the authentic protein in deletion of 39 carboxy-terminal amino acids. The amino acid sequences of the third polypeptide, HBe delta N and of HBeAgR were similar, HBe delta N differed from HBeAgR only in replacement of 3 N-terminal amino acids by 16 amino acids of beta-galactosidase. The HBcAg analogs were compared with respect to their reaction with monoclonal antibody (mAb E1A7) to the amino-terminal linear epitope of hepatitis B virus e antigen. Although able to assemble into virus-like particles, the three analogs of HBcAg, reacted differently with mAb E1A7. It was demonstrated that mAb E1A7 reacted with both native and denatured HBeAgR. HBe delta N was not recognized by mAb E1A7. In contrast, HBcAgR reacted with mAb E1A7 only when denatured. Native HBcAgR did not react with mAb E1A7 when assembled into particles. Thus evidence was obtained that the amino terminus of HBcAg is not exposed on the particle surface.

Binding, Competitive↗
PubMed1996

Design of immunogens as components of a new generation of molecular vaccines.

Three new approaches to design effective immunogens are considered. At first, we derived an expression vector from bacteriophage M13 allowing the exposure of short peptides on the virion surface. EIA demonstrates that antibodies against a recombinant phage carrying the antigenic determinant of the HIV-1 gag protein reacted with the 17-kDa core protein of the virus and also with its polyprotein precursor p55 in immunoblotting. In another approach, we chose the hepatitis B core antigen (HBcAg) particle as a vehicle for the presentation of foreign antigenic determinants to the immune system. Chimerical particles of HBcAg containing epitope of the VEE virus were obtained. A vector system for insertion of foreign antigenic determinants and production of both hybrid and wild HBcAg proteins were also obtained. The third approach relies on construction of immunogens from different T- and B-cell epitopes of the HIV-1. We suggested to construct HIV-1 vaccines in a form of the TBI (T- and B-cell epitopes containing Immunogen) with a predetermined tertiary structure, namely, a four-alpha-helix bundle. The gene of the TBI protein consisting of nine HIV-1 epitopes was synthesized and expressed in Escherichia coli cells. Mice immunized with TBI showed humoral and cellular immune responses to HIV-1. Anti-TBI antibodies displayed HIV-1 neutralizing activity. These new approaches offer promise in the development of new effective vaccines.

AIDS Vaccines↗
PubMed1993

Design of specific immunogens using filamentous phage as the carrier.

Earlier, we developed an expression vector allowing exposure of short peptides on the surface of bacteriophage M13. It was used to obtain a recombinant phage carrying an antigenic determinant of HIV1 p17 Gag protein. Immunoglobulin elicited by immunizing rabbits with the phage reacted with the 17-kDa core protein of the virus and with its polyprotein precursor, p55, on Western blots of HIV1 viral proteins. The results of present experiments may be useful in vaccine development.

Amino Acid Sequence↗
PubMed1992

Inserting foreign peptides into the major coat protein of bacteriophage M13.

Foreign DNA fragments were inserted into filamentous phage gene VIII to create hybrid B-proteins with foreign sequences in the amino terminus. The hybrid proteins are incorporated into the virions which retain viability and infectivity. Virions with hybrid B-proteins have the same contour length and the same number of B-protein molecules as virions with natural B-proteins. It was shown that for one of hybrid B-proteins the position of the processing site had changed.

Amino Acid Sequence↗