<em>Background</em><span style="font-family:;" "=""><span style="font-family:Verdana;">: In more than 50% of male subfertility, the aetiology remains unknown. Antisperm antibodies (ASA) might be involved, however the exact role of ASA in unexplained male subfertility is not clear, yet. </span><i><span style="font-family:Verdana;">Objective</span></i><span style="font-family:Verdana;">: The aim of this study was to examine 1) the prevalence of ASA in subfertile men, 2) the possible causes of the presence of ASA, and 3) the influence of ASA on sperm parameters and fertilization including assisted reproductive technologies (ART) and pregnancy outcomes. </span><i><span style="font-family:Verdana;">Study Design</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Size</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Duration: </span></i><span style="font-family:Verdana;">In this retrospective single center study, all men with semen analyses between January 2003 and December 2017 were included as well as all subfertile couples getting treatment if at least one sperm analysis showed a spermMar test ≥50%. </span><i><span style="font-family:Verdana;">Methods</span></i><span style="font-family:Verdana;">: Collected parameters were: intoxications, medication and professions that could have an adverse effect on fertility, sperm parameters, the type and amount of ART, mode of conception, pregnancy rates and outcomes. </span><i><span style="font-family:Verdana;">Results</span></i><span style="font-family:Verdana;">:</span><i> </i><span style="font-family:Verdana;">3098 semen analyses were performed. In total, 233 ASA positive men were observed, including 175 subfertile couples with an ASA positive man in the additional analyses. The prevalence of ASA in the subfertile population was 8.2%. The presence of ASA was significantly associated with the presence of oligoasthenoteratozoospermia (OAT), asthenoteratozoospermia and asthenozoospermia (p = 0