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Non-Confidential Disclosures

" A Primer-Free Method for Aptamer Selection "

PSU Invention Disclosure Number 2007-3315

Field of the Invention:

Molecular Biology

Inventors:

Weihua Pan and Gary A. Clawson

Background:

Aptamers are short polynucleotides (RNA, DNA, or other forms with modified backbones) which are capable of binding to various targets (for example, proteins) with high affinity. Aptamers are generally identified through Sequential Enrichment of Ligands by Exponential Enrichment (SELEX) procedures. These involve binding a library containing random aptamers to the target molecule, and then isolating the subset of the library which bound. This bound subset must then be reamplified by PCR, and this constitutes a “round” of selection. The process is then repeated, often 20-30 rounds, to obtain a small set of high-affinity aptamers. The reamplification is accomplished by using predetermined “flanking “ sequences, which allow the PCR amplification after each round. However, these flanking sequences are of lengths sufficient to cause binding problems, leading to a high background of identified aptamers which either do not bind well, or bind promiscuously to non-target molecules.

Invention description:

We have devised an aptamer library which allows us to separate the random aptamer sequences a large library from their flanking sequences, allowing us to bind just the random aptamer sequences to the target. After binding, the bound aptamers are reuinited with the flanking seuqneces, which allows them to be reamplified for subsequent rounds in a SELEX protocol.

Advantages:

  • Our random aptamer library and protocol can be used used for essentially any target. Other methods which have been reported are specific for given applications
  • The protocol is streamlined for ease of use
  • The protocol should always yield a very low background of non-specific binding, so that essentially all identified aptamers can be expected to have high affinity for target.

Patent status:

Patent pending.

Contact:

James F. Kolonay
Intellectual Property Office
113 Technology Center
The Pennsylvania State Univ.
University Park, PA 16802-7000
Phone: (814) 863-7070
Fax: (814) 865-3591
E-mail:jfk11@psu.edu