Temperature-responsive bioconjugates. 2. Molecular design for temperature-modulated bioseparations.
We have synthesized carboxyl semitelechelic oligo(N-isopropylacrylamide) (OIPAAm) using radical telomerization with 3-mercaptopropionic acid. This telomerization is also effective for the synthesis of carboxyl semitelechelic co-oligomers of IPAAm with butyl methacrylate (BMA) as hydrophobic or N,N-dimethylacrylamide (DMAAm) as hydrophilic comonomers. All co-oligomers are highly water-soluble at lower temperatures and exhibit phase separation with increasing temperature. Pure OIPAAm exhibits a lower critical solution temperature (LCST) at 32 degrees C, and the LCST for co-oligomers can be controlled to increase over 32 degrees C with increasing DMAAm composition and to decrease below 32 degrees C with increasing BMA composition. OIPAAm was grafted to bovine serum albumin (BSA) and bovine plasma fibrinogen (BPF) by activated ester-amine coupling. These OIPAAm-biomolecule conjugates maintain their temperature responses, are soluble in cold water, and precipitate over a range of temperatures related to oligomer content. Conjugates could be selectively precipitated and independently separated from conjugate solution mixtures with increasing temperature. In this case, the number of OIPAAm molecules attached to a conjugate affects the aggregate sizes of precipitated conjugates in mixtures. Both conjugate mixture ratios and solution concentrations influence the contamination of oligo(IPAAm-co-DMAAm)-BSA conjugates in precipitated oligo(IPAAm-co-BMA)-BPF conjugates. Furthermore, precipitated conjugates separated using centrifugation and filtration redissolve in water and maintain their biofunctionality, indicating the potential of strategy in reversible bioreactors and protein separations.
Takei
Y G
YG
Department of Chemistry, Faculty of Science and Technology, Sophia University, Tokyo, Japan.
Aoki
T
T
Sanui
K
K
Ogata
N
N
Okano
T
T
Sakurai
Y
Y
D015097:3-Mercaptopropionic Acid; D000180:Acrylic Resins / Q000737:chemistry*; D000818:Animals; D002417:Cattle; D005340:Fibrinogen / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature
D000180:Acrylic Resins; C052970:poly-N-isopropylacrylamide; D012710:Serum Albumin, Bovine; D005340:Fibrinogen; D015097:3-Mercaptopropionic Acid