Herein, we report the synthesis of a series of novel pyridyl disulfide (PDS)-functionalized statistical reactive copolymers that enable facile access to complex polymeric architectures through highly selective thioldisulfide exchange reaction with thiol-containing ligands or proteins. Functional reactive poly(N-vinyl lactam)-based copolymers including poly(N-vinylpyrrolidone-co-pyridyldisulfide ethyl methacrylate) (PVPD), poly(N-vinylpiperidone-co-pyridyl disulfide ethyl methacrylate) (PVPID), and poly(N-vinylcaprolactam-co-pyridyl disulfide ethyl methacrylate) (PVD) with PDS side groups were synthesized via reversible addition-fragmentation chain transfer (RAFT) polymerization at 60 degrees C in anisole with methyl 2- (ethoxycarbonothioylthio)propanoate as chain transfer agent. The PDS contents in the synthesized copolymers were varied from 2 to 10 mol % (as confirmed by systematical characterization with FTIR/Raman and H-1 NMR spectroscopy) using well-controlled continuous feeding method. The kinetics study suggested that copolymerizations were less favored with the enlargement of the lactam rings, indicated by lower conversions and larger dispersity indexes (D). The PDS-functionalized reactive polymers were amenable to functionalization with a variety of thiol-containing molecules, including 3-mercaptopropionic acid (3M), 2-phenylethanethiol (2P), methyl 3-mercaptopropionate (M3), 2-mercaptoethanol (2M), 2-aminoethanethiol (2A), poly(ethylene glycol) methyl ether thiol (PEG-SH), and enhanced green fluorescent protein (EGFP) via thioldisulfide exchange reaction under mild conditions, confirmed by 1H NMR and SDS-PAGE. The conversions in all cases were higher than 95%, displaying that the thioldisulfide exchange reaction to PDS groups with thiol-containing molecules is highly selective and tolerant to different ligands including amine, carboxyl, hydroxyl, phenyl, PEG and even polypeptides, providing a versatile scaffold for facile conjugation of various biological components. The contact angle measurement results and fluorescence microscopy study indicated that the reactive films based on the PDS-functionalized copolymers allowed facile, direct, and environmental-friendly surface engineering of surfaces from aqueous solution suggesting potential application in surface decoration of tissue-engineering scaffolds and medical implants. The initial cell culture experiments with HeLa cells displayed that the unmodified PVPD film was nontoxic and biocompatible while the film modified with PEG (a type of antifouling polymer) showed diminished cell attachment and growth, indicating that elegant engineering of the film surface can meet demands of particular applications.