The serum protein profiles of normal subjects, patients diagnosed with cervical cancer, and oral cancer were recorded
using High Performance Liquid Chromatography combined with Laser Induced Fluorescence detection (HPLC-LIF).
Serum protein profiles of the above three classes were tested for establishing the ability of HPLC-LIF protein profiling
technique for discrimination, using hard clustering and Fuzzy clustering methods. The clustering algorithms have quite
successfully classified the profiles as belonging to normal, cancer of cervix, and oral cancer conditions.
Protein profiles of tissue homogenates were recorded using HPLC separation and LIF detection method. The samples
were collected from volunteers with clinically normal or cervical cancer conditions. It is shown that the protein profile
can be classified as belonging to malignant or normal state by using hard and Fuzzy clustering methods. The study was
performed to test the utility of the HPLC-LIF protein profiling method for classification of tissue samples as well as to
establish a complementary method for histopathology for clinical diagnosis of the tissue as normal or malignant.
High performance liquid chromatography with high sensitivity laser-induced fluorescence detection is used to study the protein profiles of serum samples from healthy volunteers and cervical cancer subjects. The protein profiles are subjected to principal component analysis (PCA). PCA shows that the large number of chromatograms of a given class of serum samples—say normal/malignant—can be expressed in terms of a small number of factors (principal components). Three parameters—scores of the factors, squared residuals, and Mahalanobis distance—are derived from PCA. The parameters are observed to have a narrow range for protein profiles of standard calibration sets formed from groups of clinically confirmed normal/malignant classes. Limit tests using match/no match of the parameters of any test sample with parameters derived for the standard calibration sets give very good discrimination between malignant and normal samples with high sensitivity (~100%) and specificity (~94%).
HPLC combined with laser induced fluorescence provides a very sensitive method for the separation and identification
of the many proteins present in clinical samples. Protein profiles of clinical samples like Pap smear and tissue samples,
from subjects with cervical cancer and normal volunteers, were recorded using HPLC-LIF. The protein profiles were
analyzed by Principal Component Analysis (PCA). The profiles were characterized by parameters like scores of the
factors, sum of squared residuals, and Mahalanobis Distance, derived from PCA. Parameters of each sample were
compared with those of a standard set and Match/ No Match results were generated. Good discrimination between
normal and malignant samples was achieved with high sensitivity and specificity.
Proteomics based techniques are rapidly emerging as alternative techniques to conventional histo-pathological methods
for detection and diagnosis of cancers. Tumor markers are of considerable importance in the study of various cancers. A
study of various changes in the protein profile associated with breast cancer will facilitate a better understanding of the
various dynamic changes associated with the disease. In our study we have used High Performance Liquid
Chromatography coupled with highly sensitive Laser Induced Fluorescence for recording the protein profiles of breast
tissue homogenates. The protein profiles were recorded from pathologically certified normal as well as malignant breast
tissue samples. The recorded protein profiles were studied by using Principal Component Analysis. Good
discrimination of normal, benign and malignant samples was achieved in this pilot study.
Optical methods and proteomics investigations are becoming promising approaches for early detection of many diseases, which remain clinically silent for long periods. We have used efficient High Performance Liquid Chromatography (HPLC) separation combined with highly sensitive laser induced fluorescence detection of proteins present in clinical samples for diagnostic applications in cervical cancer. The protein profile and the fluorescence of individual proteins were simultaneously recorded using our HPLC-LIF system. Protein profiles (Chromatogram) of serum from normal male and female volunteers with and without tobacco habits, and malignant serum samples were studied. Protein profiles were also recorded for lysates of exfoliated cells collected from Pap smear of normal and cancer patients. The protein profile patterns were subjected to Principal component Analysis. Discrimination of normal and malignant samples were achieved with very high sensitivity and specificity.
New techniques for the early detection of cancer are fast emerging. This is essential for more effective diagnosis and control of the disease. We have used a High Performance Liquid Chromatography-Laser Induced Fluorescence (HPLCLIF) technique to record chromatograms of proteins in serum and ovarian tissue samples. The recorded chromatograms of normal, benign and malignant samples were analyzed using statistical (Principal Component Analysis) methods. It is shown that chromatograms of the samples can be classified into sets, and a model based on such a classification can be used to analyze protein profiles of test samples of serum and ovarian tissue for the detection of malignancies.
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