The synthesis of a smart streptavidin-functionalized poly(N-isopropylacrylamide) composite and its application in the separation and detection of virus nucleic acid.

A new kind of polymeric material (PNIPAAm-co-SA) was prepared by conjugating a thermosensitive polymer, Poly (N-isopropylacrylamide) (PNIPAAm) with streptavidin (SA). This smart prepared composite displayed a controllable conformation change between an expanded and a collapsed form, below or above its lower critical solution temperature (LCST). Differential scanning calorimetry (DSC) analysis demonstrated that the PNIPAAm-co-SA bioconjugate showed the same LCST as the original synthetic polymer, PNIPAAm, which was also 32°C. Based on the specific interaction between SA and biotin, a higher capture efficiency of PNIPAAm-co-SA, which was almost 100% in PBS buffer solution and above 70% in serum was obtained, respectively. And the high affinity between PNIPAAm-co-SA and biotin was still maintained after three heating cycles. Subsequently, the variola virus (small pox, VV) oligonucleotide sequence was chosen as a model to demonstrate the sensitivity of the biosensor which was fabricated based on PNIPAAm-co-SA. The biosensor exhibited the ability to separate and enrich targets from complicated system with its phase transition ability, and high sensitivity toward VV-targets were achieved. Moreover, other types of targets such as proteins and cells, could be detected by changing the biotin-captures, which indicated the broad applicability of biosensors based on this smart polymer material.