SC²: A Novel Process for Manufacturing Multipurpose High‐Density Chemical Microarrays

Abstract

Various formats of miniaturized arrays of cellulose-bound synthetic probe molecules to suit a broad range of technical and assay configurations have been produced by a novel process which we call spotting compound-support conjugates, SC 2 . This process involves parallel chemical assembly of the probe molecules by SPOT-synthesis on a special cellulose membrane, post-synthesis separation of the tethered compound spots, solubilization of the cellulose matrix and spotting of the soluble cellulose-probe conjugates onto various types of planar surfaces. The performance of such arrays are demonstrated using a set of synthetic peptide analogues, assayed with a monoclonal antibody, as well as utilizing a small molecule library of diketopiperazines assayed with fluorescently labelled calmodulin.

N-terminal myristoylation-dependent masking of neutralizing epitopes in the preS1 attachment site of hepatitis B virus

“…PreS1-antibodies appear earlier after acute infection than anti-SHBs and correlate with seroconversion to anti-HBs, and therefore, might be an early marker for recovery [9,57]. Alberti et al [1] identified antibodies against preS1 (10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21) and (21)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36) in patients with resolving acute hepatitis B but not in patients who developed chronic infection. Chronic patients had only antibodies against preS1(83-106) [1].…”

“…Peptides were synthesized by the SPOT-method [14] and the HBV scanning chip was then prepared by the SC 2 -method printed on a glass microscope slide as described [10]. The HBV chip contains 4068 spots, 12-fold repetition of the same array including celluloseconjugated biotin as a control.…”

Combinatorial Synthesis of Peptide Arrays with a Laser Printer

“…This variant of the spot synthesis is particularly useful for the production of multiple replicas of densely spaced peptide arrays. [5] Arrays with thousands of oligonucleotides per cm 2 can be synthesized by lithographic methods; [6] however, with these methods only one kind of monomer can be coupled at a time to spatially defined regions on the solid support. Thus, 20 10 coupling cycles are required to synthesize an array of peptides of 10 amino acids in length.…”

Kombinatorische Synthese von Peptidarrays mit einem Laserdrucker

“…Diese Variante der SPOT-Synthese ist insbesondere dann vorteilhaft, wenn viele Kopien von hochdichten Peptidarrays hergestellt werden sollen. [5] Mit lithographischen Methoden können mittlerweile Arrays mit tausenden von Oligonucleotiden pro cm 2 hergestellt werden. [6] Der Nachteil dieser Methoden liegt darin, dass hierbei immer nur ein Monomer pro Arbeitsschritt an den Träger gekuppelt werden kann.…”