Intraocular injection of anti-vascular endothelial growth factor (VEGF) antibodies is the first-line treatment for ocular neovascular diseases. However, the invasive nature of this administration method often reduces patient compliance and negatively affects treatment outcomes. Noninvasive formulations of anti-VEGF antibody are urgently needed, but their development remains challenging due to the complex ocular barriers. This study identified an anti-VEGF single-domain antibody (sdVE01) that is three times smaller than the commercially available ranibizumab, yet retains a comparable anti-angiogenic effect to the heavy-chain region of ranibizumab (VHHL). Additionally, four dithiolane molecules (DM) were designed to construct DM-based antibody nanoformulations, which effectively penetrate both the anterior and posterior segments of the eye. Upon eyedrop administration, DM-based antibody nanoformulations significantly inhibited the VEGF pathway and reduced neovascularization in a corneal alkali-burn rat model. Notably, the therapeutic effects of the antibody eyedrops were comparable to those of ranibizumab administered via subconjunctival injection. Overall, the dithiolane-based antibody eyedrops represent a promising noninvasive strategy for treating ocular neovascularization diseases.