Preparation of anti-morphine monoclonal antibodies with complete cross reactivity with heroin

Abstract

Abstract: Objective To prepare and characterize anti-morphine monoclonal antibodies (mAbs) with complete cross reactivity with heroin. Methods Morphine-6-succinyl was conjugated with Blue carrier (BC) in the presence of carbodi imide.BALB/c mice were immunized with the holoantigens (M-6-S-BC) and Freund adjuvant,and hybridoma was generated with mouse spleen cells and Sp2/0 myeloma cells.After 3 fusions and cloning,4 hybridoma cell lines secreting monoclonal antibodies to morphine were obtained and the mABs purified from the mouse ascites followed by characterization with enzyme-linked immunosorbent assay (ELISA). Results Four hybridoma cell lines secreting anti-morphine mAbs were established and designated as G1,G3,H4 and B1,respectively.All the mAbs belonged to IgG1 isotype and could bind to heroin better than to morphine and codeine,which were identified by competitive inhibition ELISA. Conclusion The anti-morphine mAbs can bind with the opioids,especially heroin,and can be useful for sensitive detection of opioids.

CLC Number:

R965

LI Lin¹, LIU Jie², ZHU Ping², XU Jiang-ping¹, FU Ning². Preparation of anti-morphine monoclonal antibodies with complete cross reactivity with heroin[J]. Journal of Southern Medical University, 2005, 25(07): 833-836.

References

[1] Plotkowiak Z, Popielarz-Brzezinska M, Luczak J, et al. Quality assessment of morphine hydrochloride solutions [J]. Acta Poi Pharm, 2004, 61(2): 103-6.
[2] Hida M, Mitsui T, Tsuge S, et al. Rapid and sensitive determination of morphine in street opium samples by thermal desorption gas chromatography using a microfurnace pyrolyzer [J]. J Sep Sci,2004, 27(12): 1030-2.
[3] Fernandez P, Vazquez C, Morales L, et al. Analysis of opiates, cocaine and metabolites in urine by high-performance liquid chromatography with diode array detection (HPLC-DAD)[J]. J Appl Toxicol, 2005, 25(3): 200-4.
[4] Sabzevari O, Abdi Kh, Amini M, et al. Application of a simple and sensitive GC-MS method for determination of morphine in the hair of opium abusers[J]. Anal Bioanal Chem, 2004, 379(1): 120-4.
[5] Narongchai P, Sribanditmonkol P, Thampithug S, et al. The duration time of urine morphine detection in heroin addicts by radioimmunoassay[J]. J Med Assoc Thai, 2002, 85(1): 82-6.
[6] Akbarzadeh A, Mehraby M, Zarbakhsh M, et al. Design and synthesis of a morphine-6- succinyl-bovine serum albumin hapten for vaccine development [J]. Biotechnol Appl Biochem, 1999, 30(2):139-45.
[7] Cirimele V, Etienne S, Villain M, et al. Evaluation of the One-Step ELISA kit for the detection ofbuprenorphine in urine, blood, and hair specimens[J]. Forensic Sci Int, 2004, 143(2-3): 153-6.
[8] 李志勇.细胞工程[M].北京:科学出版社,2005.160-76.
[9] 李琳,赵文忠,杨淑琴,等.抗吗啡抗体的制备及其特性的评价[J].第一军医大学学报,2004,24(6):673-6.Li L, ZhaoWZ, Yang SQ, et al. Prepatation and characterization of anti-morphine antiserum [J]. J First Mil Med Univ/Di Yi Jun Yi Da Xue Xue Bao, 2004, 24(6): 673-6.
[10] Rahbarizadeh F, Rasaee MJ, Madani R, et al. Preparation and characterization of specific and high-affinity monoclonal antibodies against morphine[J]. Hybridoma, 2000, 19(5): 413-7.
[11] Aoki K, Shikama YS, Kokado A, et al. Enzyme linked immunosorbant assay and latex agglutination inhibition reaction test for morphine in urine[J]. Forensic Sci Int, 1996, 81: 125-32.
[12] Wainer BH, Fitch FW, Rothberg RM, et al. The structure of morphine momohe misuccinate[J]. Science, 1972, 178: 647-8.