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Enzymatic cycling systems have an important requisite, namely the analyte concentration has to be well below its Michaelis-Menten-value KM . Otherwise, the reaction speed is not proportional to the analyte concentration (Bergmeyer, 1983). The combination of cycling systems gives very high amplification factors. The latter describes the ratio of amplified and non-amplified signal in the linear 50 F. Lammers · Th. Scheper Fig. 8. Amplification system of lactate oxidase (LOD), lactate dehydrogenase (LDH) and catalase (CAT) range.

1985). Mecklenburg et al. (1993) used the LOD/LDH/CAT-system for an insulin-TELISA. Here, an amplification factor of about 10 was observed. Nevertheless, the cycling system is expensive, complicated and difficult to reproduce (Lammers, 1996). Especially for TELISA procedures, an optimized substrate for peroxidase labeled antibodies was developed. The substrate (2 mmol/l H2O2 and 2 mmol/l aminoantipyrine) causes a similar TELISA sensivity to that of the LOD/LDH/CAT cycle but is much easier to use, cheaper and very reproducible (Lammers, 1996).

Danielsson B, Buelow L, Lowe CR, Satoh I, Mosbach K (1981) Anal Biochem 117:84 46. Mandenius CF, Danielsson B (1988) Meth Enzymol 137:307 47. Danielsson B (1991) In: Blum LJ, Coulet PR (eds) Biosensor Principles and Applications. Marcel Dekker, New York, p 83 48. Rank M, Gram J, Danielsson B (1992) Biosensors and Bioelectronics 7:631 49. Mattiasson B, Danielsson B, Hermannsson C, Mosbach K (1978) FEBS Lett 85(2):203 50. Preininger C, Danielsson B (1996) Analyst 121:1717 51. Satoh I (1991) Netsu Sokutei 18(2):89 52.

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