In vivo probe: the collagen hybridizing peptide – 3Helix.

The new in vivo probe sCy7.5-CHP

3Helix is our partnered collagen fiber detection company. First, we would like to introduce the new product released last year, the in vivo probe (sCy7.5-CHP).

The in vivo probe (sCy7.5-CHP) is a unique collagen hybridizing peptide (CHP) that specifically binds to unfolded collagen chains. Although the product was designed for in vivo use, binding also occurs in vitro.

The principle of the new in vivo probe

The new product works on the principle of hybridization of similar sequences. Since the CHP molecule has the same Gly-X-Y sequence as that of natural collagen, the molecule only attaches itself to damaged structures. This is because this sequence is present freely in damaged structures.

Accordingly, the process described here can be compared with the addition of two single strands of a DNA to form a double strand. Collagen normally exists as a triple helix; in the case of hybridization, a triple helix can be newly formed.

Advantages of the in vivo probe sCy7.5-CHP

A decisive advantage of the new product sCy7.5-CHP is the presence of the active monomeric form. Accordingly, a heating step is no longer necessary for activation. The possibility of direct injection into the animal is given due to the newest sequence. On the other hand, hybridization with itself fortunately does not occur.

The specific binding of the CHP molecules occurs because they bind exclusively to the damaged collagen fibers. Intact collagen structures, accordingly, have no free binding sites. Consequently, there is no non-specific binding.

In addition, the sulfonated cyanine 7.5 dye serves to label, the sCy7.5 CHP molecules, for near-infrared fluorescence (NIRF). In short, this provides another advantage, better tissue penetration, while minimizing tissue autofluorescence.

What can the new product be used for?

The sCy7.5-CHPs serve as a powerful in vivo tool for the detection of tissue damage associated with collagen structural damage. Measurements are therefore made at the molecular level.

This would include inflammation and tissue damage caused by diseases, for example. Also the tissue remodeling in developmental phases, or in the aging process, can be directly detected.

Application example: Monitoring the activity of fibrotic diseases

The new in vivo probe sCy7.5-CHP can be used to monitor fibrotic disease activity. This is evident in new work from Dr. Peter Westenskow’s lab (Roche). It shows the use of the in vivo probe in a subretinal fibrosis mouse model.

Moreover, increased collagen remodeling has been shown to correlate with progression of fibrotic disease.

Shown is the in vivo sCY7.5-CHP probe in a subretinal fibrosis mouse model.
Shown is the in vivo sCY7.5-CHP probe in a subretinal fibrosis mouse model.
Left: representative IR, FA, and cSLO images of JR5558 retinas; **P<0.01 compared with control (non-targeted) sCy7.5-CHPs.
CHPS, collagen-hybridizing peptides; cSLO, scanning laser ophthalmoscope; FA, fluorescein angiography; Ir, infrared reflectance.

Literature: Jaramillio C et al. J Pediatric Gastroenterol Nutr. 2020 Jan; 70 (1); 87-92; Jessen, H et al. Respir Res 22, 205 (2021).

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