Spider silks represent a protein-based biomaterial with outstanding mechanical properties. They consist of large and repetitive proteins composed of modular consensus motifs, showing a molecular weight from at least 250 kDa to several hundred kDa. Biotechnological production of spider silk proteins is the way to prevent limitations in all intended applications. Here, the heterologous expression of high-molecular weight spider silk proteins originated from Nephilaclavipes and spider silk-derived proteins fused to elastin-like peptides (ELPs) is presented. For that reason, three different multimerization strategies were applied. By the intein-mediated multimerization, the heterologous expression of native-sized spider silk-like proteins in planta larger than 250 kDa was shown for the first time. For further characterization, protein layers were analyzed by atomic force microscopy. In addition, the first immunological analysis of spider silk-like fusion proteins are shown. All in all, the results of this study provide an important contribution about the influence of the primary structure elements on the mechanical and biological properties.