Background: Carbohydrate-protein interactions participate in many biological functions. To characterize the binding interactions represents a longstanding challenge. Method: We developed a glycan membrane array to study the interactions of carbohydrates with lectins, proteins and viruses, including 17 lectins, four antibodies (that are specific to different Lewis antigens), one recombinant H5N1 hemagglutinin, and five influenza B clinical isolates. Results: The results were found to be comparable to previous reports, indicating the efficacy and reliability of our developed method. Conclusion: This carbohydrate membrane array represents a convenient, reliable and low-cost method to examine the carbohydrate-binding features of various proteins, high-throughput drug screening and the glycan-binding surveillance of influenza viruses.