The interest for ratiometric luminescent probes that detect and quantify a specific analyte is growing. Due to their special luminescence properties, lanthanide(III) cations offer attractive opportunities for the design of dual-color ratiometric probes. Here, we describe the design principle of hetero-bis-lanthanide peptide conjugates using native chemical ligation for a perfect control of the localization of each lanthanide cation within the molecule. Two zinc-responsive probes, r-LZF1 Tb|Cs124|Eu and r-LZF1 Eu|Cs124|Tb are described on the basis of a zinc finger peptide and two DOTA complexes of terbium and europium. Both display dual-color ratiometric emission responding to the presence of Zn 2+. Using a screening approach, anthracene was identified for the sensitization of the luminescence of two near infrared-emitting lanthanides, Yb 3+ and Nd 3+. Thus, two novel zinc-responsive hetero-bis-lanthanide probes, r-LZF3 Yb|Anthra|Nd and r-LZF3 Nd|Anthra|Yb were assembled, the former offering a neat ratiometric response to Zn 2+ with emission in the near-infrared around 1000 nm, which is unprecedented.