Effect of linker structure on surface density of aptamer monolayers and their corresponding protein binding efficiency.
A systematic study is reported on the effect of linker size and its chemical composition toward ligand binding to a surface-immobilized aptamer, measured using surface plasmon resonance. The results, using thrombin as the model system, showed that as the number of thymidine (T) units in the linker increases from 0 to 20 in four separate increments (T(0), T(5), T(10), T(20)), the surface density of the aptamer decreased linearly from approximately 25 to 12 pmol x cm(-2). The decrease in aptamer surface density occurred due to the increased size of the linker molecules. In addition, thrombin binding capacity was shown to increase as the linker length increased from 0 to 5 thymidine nucleotides and then decreased as the number of thymidine residues increased to 20 due to a balance between two different effects. The initial increase was due to increased access of thrombin to the aptamer as the aptamer was moved away from the surface. For linkers greater in length than T(5), the overall decrease in binding capacity was primarily due to a decrease in the surface density. Incorporation of a hexa(ethylene glycol) moiety into the linker did not affect the surface density but increased the amount of thrombin bound. In addition, the attachment of the linker at the 3'- versus the 5'-end of the aptamer resulted in increased aptamer surface density. However, monolayers formed with equal surface densities showed similar amounts of thrombin binding irrespective of the point of attachment.
Balamurugan
Subramanian
S
Department of Chemistry and Center for Biomodular Multi-Scale Systems, Louisiana State University, Baton Rouge, Louisiana 70803, USA.
Obubuafo
Anne
A
McCarley
Robin L
RL
Soper
Steven A
SA
Spivak
David A
DA
D052157:Aptamers, Nucleotide / Q000737:chemistry* / Q000378:metabolism*; D015374:Biosensing Techniques; D019855:Ethylene Glycol / Q000737:chemistry* / Q000378:metabolism*; D006046:Gold / Q000737:chemistry; D011485:Protein Binding; D020349:Surface Plasmon Resonance; D013499:Surface Properties; D013917:Thrombin / Q000737:chemistry / Q000378:metabolism*
D052157:Aptamers, Nucleotide; D006046:Gold; D013917:Thrombin; D019855:Ethylene Glycol