Drug-related research and development processes across the globe have been challenged due to the rise of increasingly complex diseases; however, it was also the emergence of these diseases that accelerated growth in these areas of expertise. Several molecular actions are associated with reactions caused by drugs. The kinetic and affinity information about these molecules is indispensable for the reason that merely understanding the action and affinity can help confirm whether the drug is acting on its target, and whether an interaction between two biomolecules exist.

Common techniques or systems designed for biomolecular interaction analysis include ELISA, Western Blot, Yeast-two hybrid, etc.

Nevertheless, there are some disadvantages while employing these techniques such as being time-consuming, providing inadequate amounts of information, and requiring labels…etc.

 

 

 

Traditional technology

SPR/PPR

Labeling

Needed

Not needed

Time costs

1-2 days

3-6 hrs

Limitations of affinity

N/A

1E-3 ~ 1E-14 (M)

Information

Yes or no

ka, kd, KD, etc.

 

 

Cytiva (formerly GE Life Sciences) developed the Biacore machine based on the Surface Plasmon Resonance (SPR) technique, which is a technique that can overcome apparent disadvantages and provide accurate kinetic and affinity analysis.

INB (Instant NanoBiosensors Co., Ltd.) developed the FOPPR machine based on the particle plasmon resonance (PPR) technique, which is another alternative that is similar to Biacore.

Although different techniques are available for use, the most suitable analysis platform must be chosen based on the distinct demands and situations.

 

 

Biacore™ - Surface Plasmon Resonance (SPR)

Surface plasmon resonance (SPR), combined with optical reflection systems, are employed to monitor binding events between molecules ranging from ions to viruses.

Binding of the analyte to the ligand in the mobile phase produces a gold film that induces changes in resonance. Consequently, this leads to deflection of reflected light, which indicates that there is an interaction between two molecules.

This technology provides real-time detection of binding, kinetics, affinity, specificity, and concentration, without the need for labels. Furthermore, SPR provides a much higher resolution of monitoring between biomolecules compared to traditional molecular experiments.

Overall, SPR is no doubt a vital technical service in the field of drug development.

 

spr

 

biacore

 

   

 

Demo Case:

Ligand: Anti-Beta 2-microglobulin Ab

Analyte: Beta 2-microglobulin

 

biacore

 

ka (1/Ms)

kd (1/s)

KD (M)

Rmax (RU)

tc Chi² (RU²) U-value

1835680

0.004368

2.38E-09

35.76335

50237046 0.120548 0.65

  

 

 

 

FOPPR™ - Particle Plasmon Resonance (PPR)

The unique & patented FOPPR™​​​​​​​ Technology combines optical waveguides with noble metal nanoparticles in order to provide sensitive and reliable detection of molecules. Using this technique, gold nanoparticles (AuNPs) are put in close proximity to an unclad optical fiber, and then as light propagates within the optical fiber, total internal reflection (TIR) results in an evanescent field that induces the AuNPs to undergo particle plasmon resonance (PPR).

When target-analytes interact with AuNPs - conjugated ligands, it leads to a change of refractive index near the sensing surface. Extreme sensitivity to

changes in the surrounding environment is an optical property that makes FOPPR™ an ideal technology for monitoring real-time interactions between a wide range of molecular species, including but not limited to organic drugs, oligonucleotides, proteins, and viruses.

 

 

foppr

 

Demo Case:

Ligand: Human GFAP Ab

Analyte: Human GFAP protein

 

foppr

 

Analyte

ka (1/Ms)

kd (1/s)

KD (M)

R-square

50 pg/mL – 1 ng/mL

1680000

0.00000418

2.38E-09

0.982