Preparation, Identification and Preliminary Application of the Fenvalerate Monoclonal Antibody in Six Kinds of Dark Tea.

Fenvalerate has the advantages of a broad insecticidal spectrum, high efficiency, low toxicity and low cost, and it is widely used in agriculture, especially in tea, resulting in the accumulation of fenvalerate residues in tea and the environment, posing a serious threat to human health. Therefore, the timely monitoring of fenvalerate residue dynamics is vital for ensuring the health of humans and the ecological environment, and it is necessary for establishing a fast, reliable, accurate and on-site method for detecting fenvalerate residues. Based on the methods of immunology, biochemistry and molecular biology, mammalian spleen cells, myeloma cells and mice were used as experimental materials to establish a rapid detection method of an enzyme-linked immunosorbent assay to detect the residues of fenvalerate in dark tea. Three cell lines-1B6, 2A11 and 5G2-that can stably secrete fenvalerate antibodies were obtained by McAb technology, and their sensitivities (IC50) were 36.6 ng/mL, 24.3 ng/mL and 21.7 ng/mL, respectively. The cross-reaction rates of the pyrethroid structural analogs were all below 0.6%. Six dark teas were used to detect the practical application of fenvalerate monoclonal antibodies. The sensitivity IC50 of the anti-fenvalerate McAb in PBS with 30% methanol is 29.12 ng/mL. Furthermore, a latex microsphere immunochromatographic test strip with an LOD of 10.0 ng/mL and an LDR of 18.9-357 ng/mL was preliminarily developed. A specific and sensitive monoclonal antibody for fenvalerate was successfully prepared and applied to detect fenvalerate in dark teas (Pu'er tea, Liupao tea, Fu Brick tea, Qingzhuan tea, Enshi dark tea and selenium-enriched Enshi dark tea). A latex microsphere immunochromatographic test strip was developed for the preparation of rapid detection test strips of fenvalerate.

Preparation of human monoclonal anti-C cell line from peripheral blood B lymphocytes of D--donor / 中国输血杂志

【Objective】 To establish human hybridoma cell lines, secreting monoclonal antibody against antigens of Rh blood system, from a donor with rare D--phenotype. 【Methods】 Peripheral blood B lymphocytes of an O type female donor, lacking C/c/E/e antigens on her erythrocyte, were transformed with Epstein-Barr virus (EBVs). EBVs were harvested from the cultural supernatant of B95-8 cells. The transformed lymphoblastoid cell line (LCL) secreting antibodies to C antigens were picked up and then hybridized with the myeloma SHM-D33 using electric fusion technique. Hybridoma cells were selected by HATD-Ouabain(HOTD)(Hypoxantine, Aminopterin, Thymidine, 2-Deoxycytide, and Ouabain)culture medium, microplate antibody screening and limited dilution subcloning. The monoclonal antibody was assayed by serological test and was confirmed by flow cytometry (FCM). 【Results】 From the cultural supernatant of D--peripheral blood transformed B lymphocytes, 3A6-C6, which agglutinated with R1R1(DCe/DCe)O-type RBCs but not with R2R2(DcE/DcE)O-type RBCs, was screened and preliminarily identified as anti-C. We established a hybridoma cell line secreting anti-C immunoglobulin M from B cells of D--individual successfully after hybridization with SHM-D33 myeloma cells. 【Conclusion】 The study had laid the groundwork for future research and development of human monoclonal antibodies against Rh antigens of RBC in future for diagnosis and screening purpose.

Frontiers of monoclonal antibodies: Applications in medical practices.

With the flourishing of innovation in drug discovery into a new era of personalized therapy, the use of monoclonal antibodies (mAbs) in the treatment of various ailments lies at the forefront. Major improvements in genetic sequencing and biomedical techniques as well as research into mAbs emphasize on determining new targets for advanced therapy while maximizing efficacy for clinical application. However, a balance has to be achieved concerning developing a target with low toxicity combined with high specificity and versatility, to allow a specific antibody to facilitate several biotic effects, ranging from neutralization of virus mechanisms to modulation of immune response and maintaining low global economic cost. Presently, there are approximately 30 mAbs' permitted for therapeutic use with many more being tested in clinical trials. Nevertheless, the heavy cost of mAbs' production, stowage and management as well as the subsequent hindrances to their development are outweighed by mAbs' clinical advantages. Compared to conventional drugs, since mAbs use as pharmacologic iotas have specific physical features and modes of action, they should be considered as a discrete therapeutic category. In this review, the history of mAb generation and the innovative technological applications of mAbs that has advanced in clinical practices is reviewed.

Drugs obtained by biotechnology processing / Drogas obtidas pelo processamento biotecnológico

In recent years, the number of drugs of biotechnological origin available for many different diseases has increased exponentially, including different types of cancer, diabetes mellitus, infectious diseases (e.g. AIDS Virus / HIV) as well as cardiovascular, neurological, respiratory, and autoimmune diseases, among others. The pharmaceutical industry has used different technologies to obtain new and promising active ingredients, as exemplified by the fermentation technique, recombinant DNA technique and the hybridoma technique. The expiry of the patents of the first drugs of biotechnological origin and the consequent emergence of biosimilar products, have posed various questions to health authorities worldwide regarding the definition, framework, and requirements for authorization to market such products.

Nos últimos anos, tem aumentado exponencialmente o número de fármacos de origem biotecnológica ao dispor das mais diversas patologias, entre elas destacam-se, os diferentes tipos de cancêr, as doenças infecciosas (ex. vírus AIDS/HIV), as doenças autoimunes, as doenças cardiovasculares, a Diabetes Mellitus, as doenças neurológicas, as doenças respiratórias, entre outras. A indústria farmacêutica tem recorrido a diferentes tecnologias para a obtenção de novos e promissores princípios ativos, como são exemplo a fermentação, a técnica de DNA Recombinante, a técnica de hidridoma, entre outras. A queda das patentes dos primeiros fármacos de origem biotecnológica e o consequente aparecimento dos produtos biossimilares têm colocado diferentes questões às autoridades de saúde mundiais, sobre a definição, enquadramento e exigências para a autorização de entrada no mercado deste tipo de produtos.

ESTABLISHMENT OF TWO CELL LINES SECRETING MONOCLONAL ANTIBODIES AGAINST PATHOGENIC FREE-LIVING AMOEBA IN SHANGHAI / 中国寄生虫学与寄生虫病杂志

Taking Naegleria australiensis, a species of pathogenic free-living amoeba (isolated in Shanghai in 1986), as antigen, two cell lines which provided potentially permanent source of monoclonal antibodies were established by lymphocytic hybridoma technique. The results of identification showed that: (1) the two cell lines could secret two different kinds of McAbs; (2) both of the McAbs were IgG (by gel diffusion); (3) McAbs produced in BALB/c mice were at high concentrations. One of them had a titer of ≥1 : 8 192 and the other≥l : 1 024 (by enzyme linked immunosorbent assay). A decline of the titers after purification by salting-out method was shown. One of the purified McAbs had a titer of≥1: 2 024 and the other≥l : 128.We have adopted two ways of recovering cryopreserved cells : ordinary recovering and "direct" recovering. The latter way was more practical because it could reduce the cycle of antibody production, and avoid contamination and chromosome variation. Experiments with different doses of cells revealed that, if the latter way was used, the optimal dose was 3 to 5 ? 106 cells per mouse.

ESTABLISHMENT OF HYBRIDOMA CELL LINES (G8,E10) SECRETING MONOCLONAL ANTIBODIES AGAINST HUMAN SPERMATOZOA / 中国法医学杂志

Two hybridoma cell lines (G8,E10) secreting monoclonal antibodies against human spermatozoa were established by fusing Sp2/0 mouse myeloma cells with splenocytes from Balb/c mice immunized with washed human spermatozoa. The results of the IIF, Elisa or peroxidase conjugated antibody im- munohistochemical staining demonstrated that both antibodies reacted with the spermatozoa in the human semen, testis, epididymus and vas deferens as well as the germ cells in the testis, cross reacted with the epithelial cells Of the thyroid gland, renal tubules, pancreatic ducts as well as the ilets of Langhans of the pancrease and not cross reacted with 33 normal human tissue cells, 9 different sources of body fluids and secretions as well as 8 different species of animal's spermatozoa. It is concluded that both monoclonal antibodies can be used for the species identification of semen stain in sex assault cases. The results of IIF demonstrated that each antibody bind to the localized region of the sperm cell surface. The antibody binding patterns were restricted to the acromosal cap, equatorial segment, postacromosal region, neck and the midpiece. The observed antibody binding patterns suggest that the human sperm surface is divided into a minimum of five domains. Both monoclonal antibodies were belong to IgG_1 subclass. The titre of Gs was 1∶512, while of E_(10) 1∶1024.