The smallpox vaccine based on the vaccinia virus was successfully used to 
eradicate smallpox, but although very effective, it was a very reactogenic 
vaccine and responsible for the deaths of one to two people per million 
vaccinated. Modified Vaccinia virus Ankara (MVA) is an attenuated derivative, 
also used in the smallpox eradication campaign and now being developed as a 
recombinant viral vector to produce vaccines against infectious diseases and 
cancer. MVA can encode one or more foreign antigens and thus can function as a 
multivalent vaccine. The vector can be used at biosafety level 1, has intrinsic 
adjuvant properties, and induces humoral and cellular immune responses. Many 
clinical trials of these new vaccines have been conducted, and the safety of MVA 
is now well documented. Immunogenicity is influenced by the dose and vaccination 
regimen, and information on the efficacy of MVA-vectored vaccines is now 
beginning to accumulate. In this chapter, we provide protocols for generation, 
isolation, amplification, and purification of recombinant MVA for preclinical 
and clinical evaluation.