Abstract - Expression of a Functional Recombinant Antibody Fragment in the Latex of Transgenic Hevea brasiliensis


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	Publications of MRB Latest Publications MRB Digest Journal of Rubber Research 3^rd Qtr 2008 Vol 11(3) 2^nd Qtr 2008 Vol 11(2) 1^st Qtr 2008 Vol 11(1) 4^th Qtr 2007 Vol 10(4) 3^rd Qtr 2007 Vol 10(3) 2^nd Qtr 2007 Vol 10(2) 1^st Qtr 2007 Vol 10(1) 1^st Qtr 2006 Vol 9(1) 2^nd Qtr 2006 Vol 9(2) 3^rd Qtr 2006 Vol 9(3) 4^th Qtr 2006 Vol 9(4) Submit paper/article Order Form	Abstract Expression of a Functional Recombinant Antibody Fragment in the Latex of Transgenic Hevea brasiliensis H.Y. Yeang, P. Arokiaraj, Hafsah Jaafar, Siti Arija M. Arif, S. Rajamanikam, J.L. Chan, Jafri Sharib, R. Leelavathy, Samsidar Hamzah and C.P.E. Van Der Logt Hevea genetic transformation was carried out by Agrobacterium mediation using a gene construct for an antibody single chain variable fragment (scFv) against the coat protein of the bacterium Streptococcus gordonii (Streptococcus sanguis). Gene expression was controlled by the 35S CaMV promoter and nos terminator regulatory sequences, together with the tobacco pathogenesis-related protein prla signal sequence. The gene construct also incorporated the marker nptll for transgene selection and the hydrophil-2 detection tag to monitor protein expression. Out of over 30 transgenic plants successfully transferred to the soil, 18 were checked for the presence of the scFv cDNA by PCR. All the transgenic plants were found to be positive for these inserts, whereas the four control plants tested were negative. The presence of the scFv protein in latex obtained from transgenic and control plants were analysed by ELISA. The scFv protein was detected in the latex from all the 11 transgenic plants tested while none of the four control plants tested positive. When the assay was modified so that the recombinant antibody bound to the antigen coated on the ELISA plate, six of the 11 transgenic plants were positive, indicating that these antibody fragments were functional. The recombinant proteins synthesised in the latex of the other five plants functioned poorly as antibodies. Hence, although all the 11 transgenic plants harboured the scFv gene, expression of the protein varied between the individual plants in quantity and functionality. Concentration of the scFv protein in the latex increased as the transgenic plants aged. back
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