Biomaterial Database

[Preparation of monoclonal antibodies against HA protein of subtype H1 of swine influenza virus]. 2009

Hong-tao Li, and Ming Liu, and Chun-guo Liu, and Jin-ling Du

National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin, China.

OBJECTIVE To prepare the monoclonal antibodies(mAb) against the HA protein of subtype H1 of swine influenza virus (SIV). METHODS To construct recombinant expression plasmid subtype H1 of SIV HA gene of A/Swine/Guangdong/LM/2004(H1N1).BALB/c mice of 6-8 weeks old were immunized endemically with the recombinant plasmid.The splenocytes from the immunized mice were fused with Sp2/0 myeloma cells after the last immunization. Hybridoma cells were screened by ELISA and hemagglutination inhibition (HI) tests.The activity and specificity of mAbs were evaluated by HI test and Western blot assay. RESULTS Three hybridoma cell lines secreting specific mAbs named 8C4, 8C6, and 9D6 were developed. The ELISA titer of these mAbs was 1:16 000-1:256 000, the HI titer was 1:512-1:256 000; The neutralized titer of 8C4, 8C6, and 9D6 to A/Swine/Guangdong/LM/2004(H1N1) was 10(-2.83), 10(-6.4) and 10(-5.8). Western blot analysis confirmed that mAbs only reacted with HA protein of subtype H1 of SIV. CONCLUSIONS These mAbs can be used as a useful tool to analyze the antigenic variation.They also provide the effective reagents for the rapid detection of subtype H1 of SIV.

UI	MeSH Term	Description	Entries
D007114	Immunization	Deliberate stimulation of the host's immune response. ACTIVE IMMUNIZATION involves administration of ANTIGENS or IMMUNOLOGIC ADJUVANTS. PASSIVE IMMUNIZATION involves administration of IMMUNE SERA or LYMPHOCYTES or their extracts (e.g., transfer factor, immune RNA) or transplantation of immunocompetent cell producing tissue (thymus or bone marrow).	Immunologic Stimulation,Immunostimulation,Sensitization, Immunologic,Variolation,Immunologic Sensitization,Immunological Stimulation,Sensitization, Immunological,Stimulation, Immunologic,Immunizations,Immunological Sensitization,Immunological Sensitizations,Immunological Stimulations,Sensitizations, Immunological,Stimulation, Immunological,Stimulations, Immunological,Variolations
D008807	Mice, Inbred BALB C	An inbred strain of mouse that is widely used in IMMUNOLOGY studies and cancer research.	BALB C Mice, Inbred,BALB C Mouse, Inbred,Inbred BALB C Mice,Inbred BALB C Mouse,Mice, BALB C,Mouse, BALB C,Mouse, Inbred BALB C,BALB C Mice,BALB C Mouse
D009976	Orthomyxoviridae Infections	Virus diseases caused by the ORTHOMYXOVIRIDAE.	Orthomyxovirus Infections,Infections, Orthomyxoviridae,Infections, Orthomyxovirus,Swine Influenza,Infection, Orthomyxoviridae,Infection, Orthomyxovirus,Influenza, Swine,Orthomyxoviridae Infection,Orthomyxovirus Infection
D004797	Enzyme-Linked Immunosorbent Assay	An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed.	ELISA,Assay, Enzyme-Linked Immunosorbent,Assays, Enzyme-Linked Immunosorbent,Enzyme Linked Immunosorbent Assay,Enzyme-Linked Immunosorbent Assays,Immunosorbent Assay, Enzyme-Linked,Immunosorbent Assays, Enzyme-Linked
D006385	Hemagglutination Inhibition Tests	Serologic tests in which a known quantity of antigen is added to the serum prior to the addition of a red cell suspension. Reaction result is expressed as the smallest amount of antigen which causes complete inhibition of hemagglutination.	Hemagglutination Inhibition Test,Inhibition Test, Hemagglutination,Inhibition Tests, Hemagglutination,Test, Hemagglutination Inhibition,Tests, Hemagglutination Inhibition
D006825	Hybridomas	Cells artificially created by fusion of activated lymphocytes with neoplastic cells. The resulting hybrid cells are cloned and produce pure MONOCLONAL ANTIBODIES or T-cell products, identical to those produced by the immunologically competent parent cell.	Hybridoma
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D000911	Antibodies, Monoclonal	Antibodies produced by a single clone of cells.	Monoclonal Antibodies,Monoclonal Antibody,Antibody, Monoclonal
D000918	Antibody Specificity	The property of antibodies which enables them to react with some ANTIGENIC DETERMINANTS and not with others. Specificity is dependent on chemical composition, physical forces, and molecular structure at the binding site.	Antibody Specificities,Specificities, Antibody,Specificity, Antibody
D013552	Swine	Any of various animals that constitute the family Suidae and comprise stout-bodied, short-legged omnivorous mammals with thick skin, usually covered with coarse bristles, a rather long mobile snout, and small tail. Included are the genera Babyrousa, Phacochoerus (wart hogs), and Sus, the latter containing the domestic pig (see SUS SCROFA).	Phacochoerus,Pigs,Suidae,Warthogs,Wart Hogs,Hog, Wart,Hogs, Wart,Wart Hog