Real-Time Nanoplasmonic Sensor for IgG Monitoring in Bioproduction

Abstract

Real-time monitoring of product titers during process development and production of biotherapeutics facilitate implementation of quality-by-design principles and enable rapid bioprocess decision and optimization of the production process. Conventional analytical methods are generally performed offline/at-line and, therefore, are not capable of generating real-time data. In this study, a novel fiber optical nanoplasmonic sensor technology was explored for rapid IgG titer measurements. The sensor combines localized surface plasmon resonance transduction and robust single use Protein A-modified sensor chips, housed in a flexible flow cell, for specific IgG detection. The sensor requires small sample volumes (1–150 µL) and shows a reproducibility and sensitivity comparable to Protein A/G high performance liquid chromatography-ultraviolet (G-HPLC-UV). The dynamic range of the sensor system can be tuned by varying the sample volume, which enables quantification of IgG samples ranging from 0.0015 to 10 mg/mL, without need for sample dilution. The sensor shows limited interference from the sample matrix and negligible unspecific protein binding. IgG titers can be rapidly determined in samples from filtered unpurified Chinese hamster ovary (CHO) cell cultures and show good correlation with enzyme-linked immunosorbent assay (ELISA).

Realization of user‐friendly bioanalytical tools to quantify and monitor critical components in bio‐industrial processes through conceptual design

“…Limits in the figure depend on analyte and process. As the arrows and positions of the citations [14–52] illuminate, many of the methodologies have undergone gradual improvements in performance. These achievements have moved their placement of the circular diagrams towards the intersected areas.…”

“…with biosensors based on enzyme electrodes, microgravity sensors, and thermal biosensors) but few are used in industrial practice and if so, only in process R&D. Successful commercially examples could be noted, F I G U R E 5 Analytical performance with critical limits for response time, sensitivity, selectivity, and cost effectivity. Selected methodologies are cited in the diagram: near-infrared- [15][16][17], 2D fluorescence- [18][19][20], and Raman spectroscopies [21][22][23], in-situ microscopy [24][25][26], holography [27,28], calorimetry [29,30], enzyme electrodes [31], enzyme thermistors [32][33][34], localized surface plasmon resonance [35], immuno-capacitive sensors [36][37][38][39], electrodes [40], aptamers [45], molecular imprinted polymers [47], electronic noses and tongues [49], and lateral flow sensors [52]. Optical and spectroscopic methods labelled blue, calorimetric red, methods based on biorecognition green, other methods with grey.…”

“…Limits in the figure depend on analyte and process. As the arrows and positions of the citations [ 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 ] illuminate, many of the methodologies have undergone gradual improvements in performance. These achievements have moved their placement of the circular diagrams towards the intersected areas.…”

“… Analytical performance with critical limits for response time, sensitivity, selectivity, and cost effectivity. Selected methodologies are cited in the diagram: near‐infrared‐ [ 15 , 16 , 17 ], 2D fluorescence‐ [ 18 , 19 , 20 ], and Raman spectroscopies [ 21 , 22 , 23 ], in‐situ microscopy [ 24 , 25 , 26 ], holography [ 27 , 28 ], calorimetry [ 29 , 30 ], enzyme electrodes [ 31 ], enzyme thermistors [ 32 , 33 , 34 ], localized surface plasmon resonance [ 35 ], immuno‐capacitive sensors [ 36 , 37 , 38 , 39 ], screen‐printed electrodes [ 40 ], aptamers [ 45 ], molecular imprinted polymers [ 47 ], electronic noses [ 48 ] and tongues [ 49 ], and lateral flow sensors [ 52 ]. Optical and spectroscopic methods are labelled blue, calorimetric red, methods based on biorecognition green, other methods with grey.…”

Gold Nanoparticle‐based “Mix and Measure” Fluorimetric Assays to Quantify Antibody Titer

“…A rapid, less tedious, and high throughput method to quantify mAbs is in demand to improve process monitoring and accelerate production efficiency. To address this demand, several new mAbs quantification sensors and assays have been developed, including an electrochemical sensor utilizing protein G – PEG hydrogel coated electrode that can quantify human IgG in the range of 15.6–1,000 mg/L [17] and a refractive index sensor utilizing protein A‐gold nanostructure decorated sensor chip that detects human IgG as low as 1.5 mg/L from 1 mL sample [18] . These sensors are fast with less than 5 min measurement for each test, but they are based on solid‐liquid phase binding assay and not designed for high‐throughput analysis.…”