News and Publications
The Skaggs Institute for Chemical Biology
Scientific Report 1998-1999
Artificial Antibodies, Catalytic Antibodies, and Materials Tailored Specifically
for Organic Synthesis
K.D. Janda, J.A. Ashley, O. Brümmer, R.A. Carrera, C.-W. Cho, A. Cordova,
P. Garibay,* C. Gao, H. Han, C.W. Harwig, J. Hasserodt, T. Hoffman, L. Jones,
G.-T. Kim, D.M. Kubitz, M. Kubitz, K.-J. Lee, C.-H. Lo, U.F. Mansoor, M. Matsushita,
S. Mao, G.P. McElhaney, W.A. Metz,** J. Lopez-Pellegrin, N.N. Reed, F. Sieber,
A. Simeonov, J.D. Toker, P. Toy, A.D. Wentworth, P. Wentworth, Jr., M. Wenz,
P. Wirsching, B. Zeng, B. Zhou, R.A. Lerner
* Novo Nordisk A/S and The Royal Danish School of Pharmacy, Bagsvoerd,
** Hoechst Marion Roussel, Inc., Somerville, NJ
Phage display has been intensively investigated, yet many details of the phage
particle itself have not been fully elucidated, and the possibility of alternative
display formats has not been explored. The proteins III, VI, and VIII encoded
by the phage genome have been used to display biological molecules, whereas those
encoded by genes VII and IX have not. The proteins encoded by genes VII and IX
are closely associated on the surface of filamentous bacteriophage opposite the
end harboring the widely exploited gene III protein.
We developed a phagemid format in which the heavy- and light-chain variable
regions of an antibody were fused to the amino termini of proteins encoded by
gene VII and gene IX, respectively (Fig. 1).
Significantly, the fused proteins interacted to form a functional antibody
domain on the phage surface. This result was the first time in which proteins
encoded by genes VII and IX were harnessed for displaying an antibody or any
protein on the phage surface. Our approach should be applicable to the display
of generic peptides and protein libraries that can form combinatorial heterodimeric
arrays. Consequently, the method is a major step toward the development of artificial
antibodies and the selection of novel biological activities.
The first report of esterolytic catalytic antibodies in 1986 opened the door
to use of these protein catalysts in the exploration of numerous chemical transformations.
One reaction not yet associated with antibody catalysis is a 1,3-dipolar cycloaddition.
We are exploring this reaction, which has wide application in chemical synthesis
of biologically relevant compounds. Figure 2 details the reaction scheme and
the haptenic structures we used to generate antibody catalysts. Besides obtaining
a protein catalyst to a reaction that has not been observed in Nature, we also
hope to address such issues as regioselective and enantioselective control and
antibody microenvironment and entropic effects.
With the advent of combinatorial chemistry and automated synthesis, interest
in polymer-supported reactions has been renewed. However, the polymer supports
currently used for synthesis of products other than peptides or nucleotides are
far from ideal. Determining new supports that are economically practical, have
satisfactory physical characteristics, and are inert to a diverse range of reagents
and catalysts commonly involved in a multistep combinatorial synthesis is not
a trivial matter. We have prepared a new class of resins that meet the criteria
stated (Fig. 3). The resins are based on a set of polytetrahydrofuran cross-linkers
and are promising polymer supports for solid-phase synthesis and combinatorial
Ashley, J.A., Lin, C.-H., Wirsching, P., Janda, K.D. Monitoring chemical
warfare agents: A new method for the detection of methylphosphonic acid. Angew.
Chem. Int. Ed. 38:1793, 1999.
Berg, T., Simeonov, A., Janda, K.D. A combined parallel synthesis and
screening of macrocyclic lanthanide complexes for the cleavage of phospho di-
and triesters and double-stranded DNA. J. Comb. Chem. 1:96, 1999.
Brümmer, O., Gao, C., Mao, S., Weiner, D.P., Janda, K.D. Design,
synthesis and characterization of panning reagents for the selection of metalloantibodies.
Lett. Pept. Sci. 6:295, 1999.
Gravert, D.J., Datta, A., Wentworth, P., Jr., Janda, K.D. Soluble supports
tailored for organic synthesis: Parallel polymer synthesis via sequential normal/living
free radical processes. J. Am. Chem. Soc. 120:9481, 1998.
Han, H., Yoon, J., Janda, K.D. Azatides as peptidomimetics: Solution
and liquid phase syntheses. In: Peptidomimetics Protocols. Kazmierski,
W.M. (Ed.). Humana Press, Totowa, NJ, 1999, p. 87. Methods in Molecular Medicine
Series, Vol. 23.
Han, H., Cho, C.W., Janda, K.D. A substrate-based methodology that
allows for the regioselective control of the catalytic aminohydroxylation reaction.
Chem. Eur. J. 5:1565, 1999.
Harwig, C.W., Gravert, D.J., Janda, K.D. Soluble polymers: New options
in both traditional and combinatorial synthesis. Chemtracts 12:1, 1999.
Hori, M., Gravert, D.J., Wentworth, P., Jr., Janda, K.D. Investigating
highly cross-linked macroporous resins for solid-phase synthesis. Bioorg. Med.
Chem. Lett. 8:2363, 1998.
Janda, K.D., Wentworth, A.D. A merging of chemistry and biology in
terms of antibody catalysis. Phosphorus Sulfur Silicon Relat. Elem. 144:117,
Lerner, R.A., Barbas, C.F. III, Janda, K.D. Making enzymes. Harvey
Lect. 92:1, 1998.
Paschall, C.M., Hasserodt, J., Jones, T., Lerner, R.A., Janda, K.D., Christianson,
D.V. Convergence of catalytic antibody and terpene cyclase mechanisms: Polyene
cyclization directed by carbocation-π interactions. Angew. Chem. Int. Ed.
Sieber, F., Wentworth, P., Jr., Toker, J.D., Wentworth, A.D., Metz, W.A.,
Reed, N.N., Janda, K.D. Development and application of a poly(ethylene glycol)-supported
triarylphosphine reagent: Expanding the sphere of liquid-phase organic synthesis.
J. Org. Chem. 64:5188, 1999.
Spivak, D.A., Hoffman, T.Z., Moore, A.H., Taylor, M.J., Janda, K.D. A
comparison of flexible and constrained haptens in eliciting antibody catalysts
for paraoxon hydrolysis. Bioorg. Med. Chem. 7:1145, 1999.
Taylor, M.J., Hoffman, T.Z., Yli-Kauhaluoma, J.T., Lerner, R.A., Janda,
K.D. A light-activated antibody catalyst. J. Am. Chem. Soc. 120:12783, 1998.
Wentworth, P., Jr., Janda, K.D. Antibody catalysis of phosphodiester
hydrolysis: A survey of hapten strategies. Phosphorus Sulfur Silicon Relat. Elem.
Wentworth, P., Jr., Janda, K.D. Generating and analyzing combinatorial
chemistry libraries. Curr. Opin. Biotechnol. 9:109, 1998.
Wentworth, P., Jr., Janda, K.D. Liquid-phase chemistry: Recent advances
in soluble polymer-supported catalysts, reagents and synthesis. Chem. Commun.
Yoon, J., Cho, C.-W., Han, H., Janda, K.D. Solution and soluble polymer
syntheses of 3-aminoimidazoline-2,4-diones. Chem. Commun. Issue 24:2703, 1998.
Zhao, X., Janda, K.D. Soluble polymer traceless linker investigations:
Solvent effects on the desulfonylation of polyethylene glycol (PEG) substituted
aryl alkyl sulfones with sodium amalgam. Bioorg. Med. Chem. Lett. 8:2439, 1998.
Zhao, X., Metz, W.A., Sieber, F., Janda, K.D. Expanding on the purification
methodology of polyethylene glycol (PEG) bound molecules: The synthesis of 3,5-pyrazolidinediones.
Tetrahedron Lett. 39:8433, 1998.