Real-time in situ monitoring of CRM-197 and polysaccharide conjugation reaction by fluorescence spectroscopy.

Aims: Process analytical technology (PAT) is increasingly being adopted within the pharmaceutical industry to build quality into a process. Development of PAT that provides real-time in situ analysis of critical quality attributes are highly desirable for rapid, improved process development. Conjugation of CRM-197 with pneumococcal polysaccharides to produce a desired pneumococcal conjugate vaccine is a significantly intricate process that can tremendously benefit from real-time process monitoring. Methods: In this work, a fluorescence-based PAT methodology is described to elucidate CRM-197-polysacharide conjugation kinetics in real time. Results & conclusion: In this work, a fluorescence-based PAT methodology is described to elucidate CRM-197-polysacharide conjugation kinetics in real time.

Fiber-optic surface plasmon resonance sensors in the near-infrared spectral region.

The sensitivity of fiber-optic surface plasmon resonance (SPR) sensors was improved by a factor of at least thirteen for aqueous solutions by modifying the tip geometry to allow interrogation of the surface plasmon (SP) band in the near-infrared (NIR) region. This was achieved by tuning the angle at the distal end of the SPR sensor to a dual taper of 71 degrees and 19 degrees . Using a low numerical aperture (NA) fiber-optic sensor, NA = 0.12, is necessary to obtain a functional SPR sensor working in the NIR region. Theoretical simulations using the Maxwell equations demonstrated that even higher enhancement is theoretically possible while maintaining a narrow spectral feature upon the excitation of the SP bands on gold surfaces. The manufacture of the SPR sensors yields good agreement between theoretical simulations and experimental observations. To investigate the properties of these fiber-optic SPR-NIR sensors, sucrose solutions ranging from 0 to 15 x 10(-3) in mole fraction were utilized. The increased sensitivity of the fiber-optic SPR sensors, when used to monitor biomarkers, would yield lower detection limits. The smaller sensing area, compared to planar or other fiber-optic SPR sensors, combined with an improvement of the sensitivity, would yield a dramatic reduction of the absolute amount detected by biosensors.