The invention discloses an anti-Pf332-DBL area monoclonal antibody capable of restraining the invasion of plasmodiumfalciparum, which can be specifically combined with the DBL region functional polypeptide of plasmodiumfalciparum Pf332 membrane protein, and the polypeptide has the amino acid sequence shown as SEQ ID No.1 or 2. The anti-Pf332-DBL area monoclonal antibody is prepared by fusing splenocyte and Sp2/0 cells through a DBL recombination protein immunity mouse with a HIS mark, primarily selecting by using a DBL recombination protein with a GST mark, and carrying out positive selecting by using a functional polypeptide. The anti-Pf332-DBL area monoclonal antibody has stronger function for restraining the invasion of polypide, and the function is stronger than that of an MP antibody purified from Mlli human serum infected by plasmodiumfalciparum. A monoclonal antibody which can not be specifically combined with the functional polypeptide has weaker function for restraining the invasion of polypide or has no function for restraining the invasion of polypide. The DBL region functional polypeptide can be applied to the research and development of polypeptide medicaments for treating malaria and the preparation of preventing active polypeptide vaccine.