Herein, we designed and synthesized a thermally stable carboxybetaine copolymer with a one- or three-carbon spacer between ammonium and carboxylate groups (CBMA1 and CBMA3) to create an anti-nonspecific adsorption surface with the ability to immobilize antibodies. A series of controlled poly(N,N-dimethylaminoethyl methacrylate) was successfully prepared using reversible addition-fragmentation chain-transfer (RAFT) polymerization and was derived to carboxybetaine copolymers of poly(CBMA1-co-CBMA3) [P(CBMA1/CBMA3)] with various CBMA1 contents, including the homopolymers of CBMA1 and CBMA3. Thermal stability of the carboxybetaine (co)polymers was higher than that of the carboxybetaine polymer with a two-carbon spacer (PCBMA2). Further, we also evaluated nonspecific protein adsorption in fetal bovine serum and antibody immobilization on the substrate coated with P(CBMA1/CBMA3) copolymers using surface plasmon resonance (SPR) analysis. As the CBMA1 content increased, nonspecific protein adsorption on the P(CBMA1/CBMA3) copolymer surface decreased. Similarly, the immobilization amount of the antibody decreased as the CBMA1 content increased. However, the figure of merit (FOM), defined as the ratio of the amount of antibody immobilization to that of nonspecific protein adsorption, depended on the CBMA3 content; FOM was higher when the CBMA3 content was 20-40% than those of CBMA1 and CBMA3 homopolymers. These findings will help enhance the sensitivity of the analysis using molecular interaction measurement devices, such as SPR and quartz crystal microbalance.