Proteome microarrays hold great promise for a variety of biotechnological applications including mapping protein-protein interactions, measuring drug-protein interactions and biomarker discovery. However, the need to express, purify and print thousands of human proteins at high density on a microarray substrate presents challenges which limit their wide-spread availability and use. We have developed a suite of novel technologies aimed at the production of in vitro (cell-free) expressed protein libraries, both on microarray surfaces or displayed on beads. Expressed proteins are isolated on beads using photocleavable linkers incorporated directly into the nascent protein during its synthesis using tRNA Mediated Protein Engineering (TRAMPE). In a different approach, proteins are isolated using antibodies immobilized on the beads through a photocleavable linker. Proteins can then be photo-released into solution (PC-SNAG) for conventional microarray printing. Alternatively, nascent proteins and coding agents are transferred directly from the beads to a surface to form discrete microarray spots whose dimensions match that of the beads, using a process termed photocleavable printing (PC-PRINT). PC-PRINT can also be used in conjunction with MALDI-MS by transferring proteins to MALDI substrates for downstream mass spectrometry based analyses. TRAMPE incorporated fluorescence labels also allow quantification and tracking of the proteins during their purification and deposition on the solid surface. When combined with methods to create Bead-Sorted Libraries of In Vitro Expressed Proteins (BS-LIVE-PRO), based on multiplexed solid-phase PCR and multiplexed cell-free expression reactions, this new approach provides an inexpensive method to fabricate very large scale, high density proteome microarrays (or bead displays). Furthermore, this multiplexed approach, in conjunction with rapid cell-free protein expression can facilitate the fabrication of readily customizable, on-demand microarrays of selected content. The resultant microarrays are shown to be compatible with measurements of protein-protein interaction. This work was partially funded by the SBIR grant program of the NIH.