Expression of a functional recombinant antibody fragment in the latex of transgenic Hevea brasiliensis

Yeang H. Y., . and Chan J. L., . and Van der Logt C. P. E., . and Leelavathy R., . and Rajamanikam, . and Hafsah J., . and Arokiaraj P., . and Samsidar H., . and Jafri S., . and Siti Arija M. A., . Expression of a functional recombinant antibody fragment in the latex of transgenic Hevea brasiliensis. pp. 215-225. ISSN 1511-1768

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 nptII 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 in quantity and functionality. Concentration of the scFv protein in the latex increased as the transgenic plants aged.

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Abstract

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 nptII 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 in quantity and functionality. Concentration of the scFv protein in the latex increased as the transgenic plants aged.

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Item Type:	Article
AGROVOC Term:	Hevea brasiliensis
AGROVOC Term:	Latex
AGROVOC Term:	Transgenic plants
AGROVOC Term:	Coat proteins
AGROVOC Term:	Streptococcus
AGROVOC Term:	Gene expression
AGROVOC Term:	Pathogenesis
AGROVOC Term:	Tobacco
AGROVOC Term:	Hydrophilicity
AGROVOC Term:	ELISA
Depositing User:	Ms. Suzila Mohamad Kasim
Last Modified:	24 Apr 2025 06:28
URI:	http://webagris.upm.edu.my/id/eprint/23181

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