Cyanobacteria harmful algal blooms are increasing in frequency and cyanotoxins have become an environmental and public concern in the U.S. and worldwide. In this Review, the majority of reported studies and developments of electrochemical affinity biosensors for cyanotoxins are critically reviewed and discussed. Essential background information about cyanobacterial toxins and electrochemical biosensors is combined with the rapidly moving development of electrochemical biosensors for these toxins. Current issues and future challenges for the development of useful electrochemical biosensors for cyanotoxin detection that meet the demands for applications in field freshwater samples are discussed. The major aspects of the entire review article in a prescribed sequence include (i) the state-of-the-art knowledge of the toxicity of cyanotoxins, (ii) important harmful algal bloom events, (iii) advisories, guidelines, and regulations, (iv) conventional analytical methods for determination of cyanotoxins, (v) electrochemical transduction, (vi) recognition receptors, (vii) reported electrochemical biosensors for cyanotoxins, (viii) summary of analytical performance, and (ix) recent advances and future trends. Discussion includes electrochemical techniques and devices, biomolecules with high affinity, numerous array designs, various detection approaches, and research strategies in tailoring the properties of the transducer-biomolecule interface. Scientific and engineering aspects are presented in depth. This review aims to serve as a valuable source to scientists and engineers entering the interdisciplinary field of electrochemical biosensors for detection of cyanotoxins in freshwaters.