There is increasing interest in agro-based, biodegradable and eco-friendly wood adhesives as partial replacements for petroleum-based adhesives. In this work, we studied the adhesion of cottonseed protein isolate (CPI) blended with several anionic water-soluble polymers. Anionic vinyl polymers studied included poly(acrylate), poly(acrylate-co-acrylamide), poly(vinyl sulfate), poly(vinyl sulfonate), and poly(vinyl phosphonate). Anionic polysaccharides studied included three types of carrageenan, carboxymethyl cellulose (CMC), low-methoxy pectin, alginate, and chondroitin sulfate. In general, the adhesive strength of CPI increased with the addition of anionic polymer up to a certain level and then decreased with further polymer addition. Different anionic polymers showed different enhancements. The best result for vinyl polymers was observed for the CPI/poly(vinyl sulfate) blend, which exhibited a 30% improved dry strength over CPI alone. The best results for the polysaccharides were obtained for the CPI/CMC and CPI/pectin blends, with improvements in dry adhesive strength over the CPI control of 66% and 50%, respectively. The CPI/CMC and CPI/pectin blends also showed improved hot water resistance. These findings suggest that the CPI/anionic polymer blends might be useful components in biobased wood adhesive formulations.