Design and Development of Parallel Biosensing System for

Personalized Chemotherapy Treatment

Ahmad Fairuzabadi Mohd Mansor

, Anis Nurashikin Nordin

, Kian Liang Goh

Soon Hin How

, Yumi Zuhanis Has-Yun Hashim

and Mardhiah Mohammad

Kulliyyah of Engineering, International Islamic University Malaysia (IIUM), Jalan Gombak, Kuala Lumpur, Malaysia

Kulliyyah of Medicine, IIUM Kuantan, Malaysia

International Institute for Halal Research and Training (INHART), IIUM Gombak, Malaysia

Kulliyyah of Allied Health Science, IIUM Kuantan, Malaysia

1 RESEARCH PROBLEM

Chemotherapy administration can sometimes inflict

negative side effects to the patient. The regimen or

cocktails of the drugs introduced into a patient’s body

has always needed careful consideration. Currently,

the combinations are determined by strength of the

regimen based on empirical technique; which is the

observation of the response exhibited by the patients.

However, an individual’s drug absorption rate is

influenced by many factors such as age, gender,

metabolism, disease state, organ function, drug-to-

drug interactions, genetics, and obesity.

Consequently, different patients can have different

body response towards the chemotherapy. Clinical

studies have proved that optimal treatment

effectiveness can be achieved only when the

chemotherapy treatment is individualized for each

patient (Zhang et al., 2013).

Current chemosensitivity assay such as MTT

Assay, ATP assay and molecular probes are tedious,

time consuming, labor intensive and expensive

(Kiilerich-Pedersen and Rozlosnik, 2012; Lazcka et

al., 2007). The tedious nature of these types of assays

prohibit individualized testing for patients before

chemotherapy. Therefore, there is a need for low-cost

point-of-care biosensors which can predict the

patient’s response towards different chemotherapy

regimens.

2 OUTLINE OF OBJECTIVES

Non-destructive monitoring of cell behaviors have

gained wide attention over the past decade. The

concept of Electrical Cell-Substrate Impedance

Sensing (ECIS) was pioneered by Giaever and Keese

in 1984 and evolved to be the most stable and

effective technique of measuring cultured cells on

microelectrodes with real-time impedance

monitoring (Cui et al., 2017; Hong et al., 2011).

In this research, the ECIS concept will be applied

to monitor adhesion, proliferation and death of cancer

cells in vitro due to exposure to chemotherapy drugs.

Chemosensitivity analysis will be performed using

the developed impedance biosensing system by

correlating the response of cell samples towards

several chemotherapy regimens. A comparison will

be made between tests conducted using biosensors

and the actual chemotherapy treatment prescribed to

patients.

3 STATE OF THE ART

Rapid and effective treatment of cancer is crucial to

improve patients’ quality of life and chance of

survival. Currently, cancer cell growth, apoptosis and

response to chemotherapeutic treatment involve

colorimetric assays, which require complex

laboratory equipment and extensive cell and drug

preparation. Measurements and cell preparation are

made at each endpoint, making the process labour-

intensive and high cost. As such personalized studies

on the efficacy of chemotherapeutical drugs on

patients are rarely done due to its high cost and

tedious process. Better disease-free survival rates

have been reported using neoadjuvant therapy where

treatment is given before surgery and is followed by

systemic chemotherapy (Ancona et al., 2001; Lowy

et al., 1999). In recent years, numerous efforts have

been made to develop better chemotherapeutic

regimens, resulting in improved outcomes and

prolonged survival (Sjoquist et al., 2011). To

facilitate this, there is great demand for rapid and real-

time techniques for studying cancerous cells

especially in terms of their reactions to drug and

toxins (Bramwell et al., 1997).

Mohd Mansor, A., Nordin, A., Goh, K., How, S., Hashim, Y. and Mohammad, M.

Design and Development of Parallel Biosensing System for Personalized Chemotherapy Treatment.

In Doctoral Consortium (BIOSTEC 2019), pages 15-19

Biosensors can be used to test the response of

tumours to different chemotherapy agents, similar to

how microorganisms are tested against different

antibiotics in vitro before the actual drugs are

employed in the patients. This enables an accurate

prediction as to which chemotherapy agent or

combination of agents that are likely to be successful

against the tumours without subjecting patients to

trial-and-error therapy as tumour behaviours are

unpredictable and differs from each individual

patient. This may also minimize the side effects of the

drugs as patients will not suffer unnecessarily from

ineffective treatment. Biosensors may also be used to

predict the aggressiveness of the tumour by

measuring the rate of tumour growth. This

information will be helpful to clinicians in deciding

which patients require more aggressive treatment to

prevent disease progression during treatment.

ECIS is the first impedance-based technique for

studies of quantifying cell behaviours. In ECIS, a

small gold electrode (250µm) is immersed in culture

medium at the bottom of the tissue culture wells.

Electrode surface is pre-coated with certain proteins

to enhance cell adhesion with the electrode. Two

electrodes, working and counter electrodes, exist in

the system. A relatively small circular gold electrode

behaves as a working electrode as compared to the

larger counter electrode at the bottom. In ECIS

biosensor systems, an AC signal with 1V amplitude

is applied through a 1MΩ series resistor at 4 kHz

frequency. The voltage across the electrodes is

measured using an amplifier (Luong et al., 2001).

Various cellular study using ECIS has been reported

such as in monitoring growth, proliferation and

differentiation of cells, cell migration and

cytotoxicity (Cui et al., 2017; Anh-Nguyen et al.,

2016; Sun et al., 2013; Mansor and Nordin, 2018)

4 METHODOLOGY

There are several aspects that need to be considered

when developing a biosensing system to predict the

outcome of chemotherapy. ECIS is the gold standard

for monitoring cellular interaction towards drugs

exposure using impedance biosensing technique. This

technique, however, requires expensive equipment

and electrodes to be implemented in high throughput

testing (HTT). By leveraging the cheap mass

fabrication costs of printed circuit board (PCB) in the

electronics industry, we propose as an alternative

Lab-on-Chip the Lab-on-PCB as a single use,

disposable biosensor. Optimization of electrode

configuration will be done analytically and

experimentally to find the best design. A portable and

wireless impedance data acquisition system will also

be developed by embedding commercialized

AD5933 Integrated Circuit (IC) impedance converter

IC with microcontroller. Finally, chemosensitivity

analysis will be performed using the developed

impedance biosensing system by correlating the

response of cell samples toward several

chemotherapy regimen tested using biosensor and the

actual chemotherapy outcome of the patients.

4.1 Design of Electrodes

The extensive research efforts in lab-on-a-chip (LoC)

in biomedical field have shown the advantages and

feasibility of the devices in real-life application.

However, despite the said advantages, LoC has less

commercialization potential due to expensive setup

for mass-manufacturing (Moschou and Tserepi,

2017). Recently technology of lab-on-PCB (lab-on-

printed circuit board) has re-emerged as a potential

alternative to LoC in biomedical field. PCB is widely

used in the electronics industry, thus established

manufacturing companies for fabrication process can

easily be found. The biosensor proposed is a thin film

nickel-gold finishing plated on copper electrodes

through electroplating. Previous study found that the

gold-plated PCB was biocompatible with human

K562 cells (Mazzuferi et al., 2010).

Optimization of electrodes will be done to

determine the highest electric field generated on the

electrode surface. This proposed design will also be

compared with conventional Interdigitated Electrodes

(IDEs) configuration to come out with the best

design. Width (w), spacing (s) and length (l) will be

varied and simulated using COMSOL Multiphysics

to find the optimum design. The design of the IDEs is

optimized to maximize sensitivity towards changes in

the cells using ECIS technique. the IDEs geometry

will be optimized such that both the cut-off frequency

of the interfacial impedance and the solution

resistance are minimized. This allows the highest

electric field to be generated by the IDEs.

4.2 Design of Data Acquisition System

Although many researches have been conducted

using the impedance monitoring technique, most

work rely on impedance spectroscopy measurement

using traditional impedance measurement

instruments such as HP 4284A precision LCR meter

(Zou et al., 2007), HP 4194A Impedance/Gain-Phase

Analyzer (Webster et al., 2009), Agilent 4294A

Impedance Analyzer (Price et al., 2009). However,

DCBIOSTEC 2019 - Doctoral Consortium on Biomedical Engineering Systems and Technologies

these traditional instruments are mostly bulky,

expensive and are difficult to be used in a portable

environment.

Intensive research has been growing to construct

a miniaturized module to replace these

instrumentations for point of care setting. The first

commercially available impedance network analyzer

implemented as a single integrated circuit (IC) was

designed and introduced by Analog Device Inc that

combines a frequency generator with a 12-bit,

1MSPS (sampling per second), analog-to-digital

converter (ADC). In this research, AD5933 will be

embedded with microcontroller unit (MCU) to make

a portable wireless data acquisition system for

impedance monitoring of the cells.

4.3 Biological Test

4.3.1 A549 Cell Lines Test

The first biological study using A549 lung cell lines

will be conducted to test the performance and

biocompatibility of the sensor. This is to come out

with the optimal electrode configuration and to ensure

the sensors are non-toxic and suitable for monitoring

cellular activities in vitro.

Electrode surface will be coated with extracellular

matrix (ECM) coating such as collagen type I to

promote cellular adhesion on to the surface of the

electrodes.

4.3.2 Chemosensitivity Test

Once the design is fixed, the chemosensitivity test

will be performed using primary lung cancer cells.

Samples will be taken from biopsy of the patients and

will be cultured in the lab until it reached suitable

passage for biosensor testing. Targeted samples are

between 10-30 different samples, as an essential

standard for pilot study of clinical device.

Chemosensitivity response of conventional

chemotherapy drugs used in treating lung cancer will

be tested against the cultured cells on the sensor, to

predict the response of each drugs on the cells.

Correlation between the sensors’ results with the

actual chemotherapy response of the patients will be

made to analyse the significance of the results

predicted by the sensors.

5 EXPECTED OUTCOME

The expected outcome of this project would be a

chemosensitivity device that is able to predict the

response of chemosensitivity based on in vitro cell

culture using the ECIS techniques. The general

system architecture is shown in Figure 1 below.

Figure 1: Overall system architecture for personalized

chemotherapy response.

There are three main parts in the system. The first

part is the impedance biosensor, which will be

fabricated using gold-plated PCB. The electrodes will

be applied with low alternating voltage at 10kHz

frequency for monitoring the cellular behaviour based

on cellular adhesion on the electrodes.

The second part is the portable impedance

measurement system, which mainly consist of

AD5933 impedance analyser IC which will be

controlled by MCU together with the Analog Front

End (AFE) as the signal conditioning circuit for

controlling current exposure to the cells on the

electrodes.

The last part is the wireless data acquisition

system, which is also controlled by the MCU

connected with the network. All raw data measured

by the system will be transferred and stored in a cloud

server and can be remotely access from other

location.

The system will be embedded in a small device

packaging to make it suitable to be placed inside the

incubator during testing. Cellular behaviour of the

cells can continuously be monitored for several days

Design and Development of Parallel Biosensing System for Personalized Chemotherapy Treatment

to show the response of chemotherapy drugs on the

cancer cells.

6 STAGE OF THE RESEARCH

This research has arrived at the stage of development

of data acquisition system and sensor validation.

Electrode design was analytically optimized to obtain

the best configuration for the sensor. Simulation

using COMSOL was done such as shown in Figure 2.

Optimization of electrode configuration was done in

term of finding lower cut-off frequency and

minimizing bulk resistance by varying width of

electrodes (W), spacing between electrodes (S) and

total number of electrodes (N). After simulation, the

design was sent out for fabrication using gold coated

PCB technique. Prototype sensor is shown in Figure

Figure 2: COMSOL Simulation of optimizing the

parameter for the electrode configuration. Simulation

suggested N=18 is the optimal number of electrodes for

having saturated average electric field on electrode’s

surface.

As of the current stage, sensors are tested with

A549 lung cancer cell lines to optimize cell seeding,

coating concentration and to check for toxicity of the

biomaterial. Next step involves development of

wireless data acquisition system and establishment of

primary lung cancer cell culture protocol before

chemosensitivity testing using biosensor. Figure 4

shows the flows of expected outcome of the research.

Figure 3: Fabricated sensor with gold coated surface

finishing PCB.

Figure 4: Flows of expected outcome of the research.

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Design and Development of Parallel Biosensing System for Personalized Chemotherapy Treatment