Serological characteristics and gene mutation analysis of Para-Bombay blood group / 军事医学

Objective To identify the Para-Bombay blood group on the basis of its serological characteristics .Methods ABO blood typing , H antigen detection , absorption and elution test , and saliva neutralization test were conducted for serological identification of ABO blood group .PCR-SSP was used to sequence FUT1 and FUT2 genes.Results Results of ABO genotyping of eight individuals of the Para-Bombay blood group were consistent with results of their serological blood typing.Among these cases, there were 3 cases of Amh,4 cases of Bmh,and 1 case of Abmh.The results of their FUT1 genotyping were h1h1 in 3 cases, h2h2 in 2 cases and h1h2 in 3 cases.Conclusion The differentce of agglutination intensity between Ac and Bc in reverse ABO blood typing and abnormal Oc agglutination is of greet significance for Para -Bombay blood group.

Optimized selection of anti-tumor recombinant antibodies from phage libraries on intact cells.

Generation of human recombinant antibody libraries displayed on the surface of the filamentous phage and selection of specific antibodies against desirable targets allows production of fully human antibodies usable for repeated administration in humans. Various lymphoid tissues from immunized donors, such as lymph nodes or peripheral blood lymphocytes from individuals with tumor or lymphocytes infiltrating tumor masses may serve as a source of specific anti-tumor antibody repertoire for generation of tumor-focused phage display libraries. In the case of lack of tumor-associated antigens in the purified form, high affinity anti-tumor antibodies can be isolated through library panning on whole cells expressing these antigens. However, affinity selection against cell surface specific antigens within highly heterogeneous population of molecules is not a very efficient process that often results in the selection of unspecific antibodies or antibodies against intracellular antigens that are generally useless for targeted immunotherapy. In this work, we developed a new cell-based antibody selection protocol that, by eliminating the contamination of dead cells from the cell suspension, dramatically improves the selection frequency of anti-tumor antibodies recognizing cell surface antigens.

Rapid Serological Identification for Monophasic Salmonella Serovars with a hin Gene: Specific Polymerase Chain Reaction

Salmonellosis is one of the most common food born diseases in Korea. However, it takes more than 8 days and many expensive antiserums are used for the identification of Salmonella serovars since the microorganism easily undergoes phase variation. According to the data that 65.5% of Salmonella isolates in 2000~2004 year had monophasic flagella, we have developed a rapid serological identification method using a hin gene-specific polymerase chain reaction (PCR) for monophasic Salmonella isolates that does not require the time-consuming phase conversion experiments. Using our new method, 'hin specific PCR-based serological test', we could identify serovars of monophasic Salmonella in 4 days. For the purpose of rapid identification of salmonella serovars collected from outbreaks and sporadic cases, hin specific PCR-based serological tests will be a fast and efficient method.

Search colorectal cancer antigen genes by screening the colorectal tissue cDNA phage expression libraries with SEREX / 解放军医学杂志

Objective Identification of colorectal cancer specific antigens from cDNA phage expression library by recombinant expression cloning (SEREX). Methods The cDNA phage expression library derived from colorectal cancer tissue was constructed using the SMART (Switching Mechanism at 5'end of RNA Transcript) techniques. The cDNA phage expression library was screened with SEREX (serological identification of antigen by recombinant cDNA expression libraries), the positive clones encoding antigenic genes were obtained after immunoscreening, and the nucleotide sequences of cDNA inserts were determined and analyzed with BLST software in GenBank, and the antigenic gene function was analyzed by bioinformatics. Results The cDNA phage expression library derived from colorectal cancer tissues was successfully constructed. The primary library consisted of 2.39?10 6 recombinants, and the recombinant rate was more than 97.5%. The titer of the amplified cDNA phage expression library was 4.1?10 10pfu/ml, and the size of inserted cDNA varied from 0.5 to 4.0kb. Sixteen positive clones encoding antigenic genes were obtained after immunoscreening, and results showed that 16 reactive clones were derived from 12 different genes. Ten of 12 genes were highly homologous to the genes known in GenBank, such as IFITM1, CD24, Survivin, KLK6, et al. Two expressed sequence tags (ESTs) were not found in GenBank. The antigenic genes included structural gene, regulation gene and metabolizing gene. Conclusion The tumor-associated antigen genes and the two ESTs were obtained by SEREX were worthy of further study on their structures and functions.