The influence of hapten size and hydrophobicity on the catalytic activity of elicited polyclonal antibodies

Journal of the American Chemical Society

Volumn 118, Issue 1, 1996, Pages 251-252

  Wallace, Michael B ^a   Iverson, Brent L ^a  

^a UNIVERSITY OF TEXAS AT AUSTIN   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

EID: 0000704799     PISSN: 00027863     EISSN: None     Source Type: Journal    
DOI: 10.1021/ja9524311     Document Type: Article

References (26)

1. Lemer, R. A., Benkovic, S. J.; Schultz, P. G. Science 1991, 252, 659-667.
2. Schultz, P. G.; Lerner, R. A. Acc. Chem. Res. 1993, 26, 391-395.
3. Stewart, J. D.; Liotta, L. J.; Benkovic, S. J. Acc. Chem. Res. 1993, 26, 396-404.
4. Tramontano, A.; Janda, K.; Lerner, R. A. Science 1986, 234, 1566-1570.
5. Pollack, S. J.; Jacobs, J. W.; Schultz, P. G. Science 1986, 234, 1570-1573.
6. Stephens, D. B ; Iverson, B. L. Biochem. Biophys. Res. Commun. 1993, 192, 1439-1444.
7. Wilmore, B. H.; Iverson, B. L. J. Am. Chem. Soc. 1994, 116, 2181-2182.
8. Stephens, D. B.; Wilmore, B. H ; Iverson, B. L. Bioorg. Med. Chem. 1994, 2, 653-658.
9. Iverson, B. L. CHEMTECH 1995, 25, 17-21.
10. Gallacher, G.; Jackson, C. J.; Searcey, M.; Badman, G. T.; Goel, R ; Topham, C. M.; Mellor, G. W.; Brockelhurst, K. Biochem. J. 1991, 279, 871-881.
11. Gallacher, G.; Searcey, M.; Jackson. C. J.; Goel, R.; Brockelhurst, K. Biochem. J. 1992, 284 , 675-680.
12. Gallacher, G.; Jackson, C. J.; Searcey, M.; Goel, R.; Topham, C. M.; Mellor, G. W.; Smith, C. Z.; Brockelhurst, K. Eur. J. Biochem. 1993, 214. 197-207.
13. Suzuki, H.; Sasaki, K.; Mukouyama, E. B.; Kawamura-Konishi, Y.; Wada, Y.; Ono, M. Biochem. Mol. Biol. Int. 1994, 34, 27-35.
14. 15
15. These values were determined spectrophotometrically using the method described in the following: Habeeb, A. F. S. A Anal. Biochem. 1966, 14, 328-332.
16. 7-9 Ten mice were used for each hapten to ensure that enough polyclonal antibody, ∼10 mg, was generated to carry out the required number of catalytic assays.
17. A number of pieces of evidence verified that the observed catalysis was due to the elicited antibodies and not contaminating enzymes. First, the control polychlonal antibodies isolated after five immunizations of 10 mice with underivatized BSA displayed no catalytic activity with any of the substrates. In addition, the observed substrate selectivity is consistent with the expected programmability of antibody catalysis. Finally, the catalytic activity that was observed was quantitatively inhibited in a specific way with hapten. Haptens 1 and 2 were analyzed for their ability to inhibit catalysis by both the anti-1 and anti-2 polyclonal samples reacting with their directly analogous substrates. Only the same hapten used to elicit the antibodies effectively inhibited the reaction when as much as a 5-fold excess was used.
18. Stahl, M.; Goldie, B ; Bourne, S. P.; Thomas, N. R. J. Am. Chem. Soc. 1995, 117, 5164-5165.
19. Jacobs, J.; Schultz, P. G.; Sugasawara, R.; Powell, M. J. Am. Chem. Soc. 1987, 109, 2174-2176.
20. Tawfik, D. S.; Zemel, R. R.; Arad-Yellin, R.; Green, B.; Eshhar, Z. Biochemistry 1990, 29, 9916-9921.
21. In fact, the vast majority of catalytic antibody studies have utilized haptens that fall in the size range represented by 1 and 2.
22. Similar ideas have been expressed in work involving minimal or attenuated haptens or substrates. See. for example, ref 20 above and the following: Janda, K. J.; Benkovic, S. J.; McLeod, D. A.; Schloeder, D. M.; Lerner, R A. Tetrahedron 1991, 47, 2503-2506.
23. Absolute affinities can sometimes be underestimated by the competition ELISA method used here. On the other hand, the relative order of determined affinities is accurate. Butler, J. E. In The Structure of Antigens, Vol. I; Van Regenmortel, M. H. V., Ed . CRC Press: Boca Raton, FL, 1992, pp 209-259.
24. Analysis of the antibodies elicited with hapten 5 was complicated by the lower solubilities of substrate 10 and especially the alcohol 15. Nevertheless, it is significant that the antibodies elicited by hapten 5 failed to exhibit catalytic activity, so these data were included in the table.
25. Davies, D. R.; Padlan, E. A. Annu Rev. Biochem 1990, 59, 439-473.
26. Of course, other size-dependent molecular properties of the haptens, such as conformational differences, cannot be ruled out as contributing to the catalytic activity, or lack thereof, of elicited antibodies.