Sains Malaysiana 50(11)(2021): 3297-3302

http://doi.org/10.17576/jsm-2021-5011-13

Trichloroacetic Acid/Acetone Precipitation Method to Optimize Canine Synovial Fluid for One and Two-Dimensional Gel Electrophoresis Studies

(Kaedah Pemendakan Asid Trikloroasetik/Aseton untuk Mengoptimumkan Cecair Sinovial Kanin dalam Kajian Elektroforesis Gel Satu dan Dua Dimensi)

WEI MIAO TAN¹, SENG FONG LAU¹*, SHARIFAH ZAKIAH SYED SULAIMAN¹, NOR SHAHIRA SOLEHAH UMRAN¹, MOKRISH AJAT¹, ROZAIHAN MANSOR¹, PUTERI AZAZIAH MEGAT ABD-RANI¹, INTAN NURFATIHA SHAFIE¹, ANGELA MIN HWEI NG² & NORASFALIZA BINTI RAHMAD³

¹Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43300 UPM Serdang, Selangor Darul Ehsan, Malaysia

²Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

³Agro-Biotechnology Institute Malaysia, National Institute of Biotechnology Malaysia, 43300 Serdang, Selangor Darul Ehsan, Malaysia

Diserahkan: 7 Disember 2020/Diterima: 28 Februari 2021

ABSTRACT

The challenge associated with the use of synovial fluid for osteoarthritic proteome studies is the optimization step, which involves the depletion of high abundant proteins from the samples. The objective of this study was to develop a cost efficient and effective method to remove albumin from canine synovial fluid for proteome studies. Pooled synovial fluid samples were obtained from clinically healthy dogs (n=5), with no radiographic features of osteoarthritis. The acetone precipitation method and 10% w/v of trichloroacetic acid/acetone were chosen to deplete the albumin from canine synovial fluid and the outcome from the different methods were compared using one dimensional and two-dimensional gel electrophoresis studies. The results showed that the 10% w/v TCA/acetone precipitation method removed highly abundant proteins from synovial fluid for gel electrophoresis studies compared to the acetone precipitation method. The 10% w/v TCA/acetone precipitation method provides an effective method to remove albumin from the synovial fluid using gel electrophoresis, especially two-dimensional gel electrophoresis. The accessibility and cost of TCA and acetone make this method of precipitation a simple and cost-effective technique in preparing a canine synovial fluid for two-dimensional gel electrophoresis analysis.

Keywords: Albumin; biomarkers; gel electrophoresis; osteoarthritis; synovial fluid

ABSTRAK

Satu cabaran dalam kajian proteomik berkaitan osteoartritis dengan menggunakan cecair sinovium adalah langkah pengoptimuman yang melibatkan pengurangan protein berkelimpahan tinggi daripada sampel. Objektif penyelidikan ini adalah untuk mengkaji kaedah yang lebih berkesan dan jimat bagi mengeluarkan albumin daripada cecair sinovium kanin untuk kajian proteomik. Sampel cecair sinovium yang dikumpulkan telah diperoleh daripada anjing yang dikenal pasti sihat secara klinikal (n=5), serta tidak mempunyai sebarang ciri-ciri osteoartritis berdasarkan keputusan radiografi. Kaedah mendakan aseton dan 10% w/v asid trikloroasetik/aseton telah dipilih untuk mengeluarkan albumin daripada sampel cecair sinovium kanin dan hasil daripada kaedah berbeza ini telah dibandingkan dengan menggunakan kajian elektroforesis gel satu dimensi dan dua dimensi. Hasil kajian menunjukkan kaedah 10% w/v TCA/aseton berjaya mengeluarkan protein berkelimpahan tinggi daripada elektroforesis gel berbanding kaedah mendakan aseton. Kaedah 10% w/v TCA/aseton telah berjaya menghasilkan satu kaedah yang berkesan untuk mengeluarkan albumin daripada cecair sinovium, terutamanya untuk kajian elektroforesis gel dua dimensi. Kebolehcapaian dan kos TCA dan aseton membuatkan kaedah mendakan ini lebih ringkas dan jimat dalam penyediaan cecair sinovium kanin untuk analisis elektroforesis gel dua dimensi.

Kata kunci: Albumin; cecair sinovium; elektroforesis gel; osteoartritis; penanda biologi

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*Pengarang untuk surat-menyurat; email: lausengfong@upm.edu.my