Anaerobic ammonium oxidation (Anammox) process is a high-rate nitrogen removal technology that has been applied in sludge dewatering effluents treatment with nitrogen removal rate as high as 9.5 kg/(m³·d). However, due to the slow growth rate of the autotrophic Anammox bacteria and the susceptivity to environmental conditions, the start-up of Anammox process is very long; the operation is unstable; and the nitrogen removal from organic-containing and/or toxicant-containing ammonium-rich wastewaters using Anammox process becomes difficult. Thus, the application of this high-rate process is significantly limited. In this paper, a newly-developed Anammox process with sequential biocatalyst (Anammox biomass) addition was established based on the procedure in fermentation engineering. We introduced the Anammox process with sequential biocatalyst addition on start-up, stable operation and the treatment of organic-containing and toxicant-containing ammonium-rich wastewaters. Results show that supplementing high-activity Anammox biomass into reactors will increase the amount of as well as the ratio of Anammox bacteria. Thus, the innovative Anammox process with sequential biocatalyst addition not only accelerates the start-up course, but also enhances the stability of Anammox process. Furthermore, it overcomes the drawbacks of wastewaters containing high organic content and toxic substances. Therefore, the application of Anammox process may be further enlarged.