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12
CONTENTS
0-8493-1141-1/02/$0.00+$1.50
© 2002 by CRC Press LLC
Quantification of Cell-
Penetrating Peptides
and Their Cargoes
Maria Lindgren, Mattias Hällbrink,
and Ülo Langel
12.1 Introduction ..................................................................................................263
12.2 Quantification Methods................................................................................264
12.2.1 Studies of Cell-Penetrating Peptides Using Reporter Groups ........266
12.2.1.1 Radiolabeling ....................................................................266
12.2.1.2 Biotinylation and Cell-ELISA..........................................267
12.2.1.3 Fluorescence Microscopy, Spectrophotometer,
and FACS..........................................................................267
12.2.1.4 HPLC Detection................................................................269
12.2.1.5 Immunodetection ..............................................................270
12.2.1.6 Detection of Enzyme Activity ..........................................270
12.2.1.7 Fluorogenic Construct Assay............................................270
12.2.2 Quantification of Biological Activity of Cargo Delivered
by CPPs............................................................................................272
12.2.2.1 Detection of Bioactivity of Peptide Cargo.......................272
12.2.2.2 Detection of DNA/PNA Antisense Effects ......................273
12.2.2.3 Quantification of Drug Activity........................................273
12.3 Summary ......................................................................................................274
References..............................................................................................................274
12.1 INTRODUCTION
The detection of intracellular cell-penetrating peptides (CPPs) poses several
obstacles — for example, the difficulty of distinguishing membrane-bound CPP from
intracellularly localized CPP. In this chapter we focus on methods used in quantifying
the efficiency of CPP internalization, as well as the quantification of cargoes delivered
by CPPs.
For efficient detection of a translocated CPP, a reporter group is generally
attached to the active amino acid sequence; this group could also be called cargo
263