Peptide-Based Drug Design

48 Winkler and Campbell
Table 1
Recommended Literature
Description Ref.
First description of the spot technique in detail (1)
Protocols for spot synthesis using active esters (2)
Automated spot synthesis using HOBt/DIC activation (3)
Comparison of OPfp and HOBt/DIC activation (4)
Investigations on the quality of spot syntheses (5)
Spot synthesis and applications (6)
Overview over spot synthesis of small organic molecules (7)
Characterization of some useful types of filter paper and description of some (8)
linkers
Modifications of cellulose and polypropylene membranes (9)
Short review of applications (10)
Review of applications (11)
Detailed review regarding synthesis and application of spot synthesis (12)
Detection of weak binding by electrochemical blotting to PVDF membranes (13)
Spot synthesis and screening on living cells (14)
method on cellulose membranes (1). One reason for this is that cellulose-based
filter paper is available in virtually every laboratory, inexpensive, and easy to
handle. In this publication we offer a short introduction of this synthesis and
screening technique and describe some common useful probing methods. Due
to the broad range of possible applications for probing, the described protocols
are only recommendations (see Table 1) (15).
2. Materials
2.1. Spot Synthesis of Macroarrays and Screening Techniques
1. Solvents: N,N´-dimethylformamide (DMF; VWR), methanol or ethanol (VWR),
N-methylpyrrolidone (NMP; Fluka), diethyl ether (VWR), dichloromethane
(methylene chloride, DCM; VWR). Solvents for washing steps should be of at
least ACS quality, whereas solvents for dissolving reagents must be amine and
water free. Organic solvents (except for methanol and ethanol) should be stored in
the dark.
2. Homemade membranes are prepared from filter paper Whatman 50, Whatman
540, or Chr1 (Whatman, Maidstone, UK) (16,17). Ready-to-use membranes are
available from AIMS Scientific (Braunschweig, Germany).