African swine fever is an economically important haemorrhagic disease of domestic pigs, caused by the large double stranded DNA virus African swine fever virus (ASFV)1. Despite being shown to have some protective effects, early research quickly ruled out the possibility of ASFV-specific antibodies having neutralising capabilities2. Key neutralisation studies have all used non-purified tissue culture supernatant as a source of virus, however recent work has shown that this can trigger macropinocytosis in host macrophages, allowing ASFV to bypass clathrin-mediated endocytosis (CME)3. This project aims to re-investigate neutralising antibody activity to determine whether this has affected our understanding of neutralisation. ASFV-specific antibodies targeting CME may be present, and may have protective effects against infection, however the effects of these antibodies might not have been detected in previous studies. This work will investigate antibodies that can recognise purified virus and the role they may play in preventing the initial stages of host cell entry. This work will also investigate other protective effects of ASFV-specific antibodies, such as antibody- and complement-mediated cytolysis. By using purified virus we can ensure we are focussing on the effects of antibodies specific to ASFV without these actions being masked by additional entry pathways, and through the use of CME and macropinocytosis inhibitors we can distinguish between pathways targeted by these antibodies to identify potential neutralising activity. This will further our understanding of the role ASFV-specific antibodies play in the response to ASFV infection, and may impact current vaccine development strategies. 1. Escribano JM, Galindo I, Alonso C. Antibody-mediated neutralization of African swine fever virus: Myths and facts. Virus Research. 2013 Apr 1;173(1):101–9. 2. Galindo I, Alonso C. African Swine Fever Virus: A Review. Viruses. 2017 May;9(5):103. 3. Hernáez B, Guerra M, Salas ML, Andrés G. African Swine Fever Virus Undergoes Outer Envelope Disruption, Capsid Disassembly and Inner Envelope Fusion before Core Release from Multivesicular Endosomes. PLOS Pathogens. 2016 Apr 25;12(4):e1005595.