A novel method to generate aptamers against recombinant targets displayed on the cell surface.
Isolation of anti-protein aptamers is typically carried out against purified recombinant protein. Thus, certain cell surface proteins including pharmacologically important receptors do not become targets of aptamers due to the difficulties encountered in their purification. In order to overcome this limitation, we developed a new SELEX procedure in which cells displaying the target protein on the cell surface are used, thereby precluding the need for target purification. As a model experiment, CHO cells expressing human transforming growth factor-beta type III receptor (TbRIII) were used as the target for SELEX, resulting in the successful isolation of a specific, high-affinity aptamer.
Ohuchi
Shoji P
SP
Department of Basic Medical Sciences, IMSUT, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan.
Ohtsu
Takashi
T
Nakamura
Yoshikazu
Y
D000818:Animals; D052157:Aptamers, Nucleotide / Q000378:metabolism; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D011509:Proteoglycans / Q000235:genetics / Q000378:metabolism; D018125:Receptors, Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism*; D052156:SELEX Aptamer Technique
D052157:Aptamers, Nucleotide; D008565:Membrane Proteins; D011509:Proteoglycans; D018125:Receptors, Transforming Growth Factor beta; D011994:Recombinant Proteins; C071736:betaglycan