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Profiling methodology in the synovial fluid of horse, dog and pig: Study of the peptidic profile with ZipTip® C18

Álvaro Fernández Manzano defended this presentation at the Faculty of Veterinary Medicine of the Complutense University of Madrid in order to obtain the Master of Research

September 17th, 2014

Our study focuses on establishing a specific and standardized Profiling protocol for the analysis of synovial fluid in dogs, horses and pigs, using the ultrafleXtreme™ MALDI - TOF / TOF (Bruker Daltonics) for this purpose. We used synovial fluid of 11 horses (n = 7 control and n = 4 cases – that included diseases such as osteochondritis dissecans, traumatic injuries, osteoarthritis and septic arthritis), 9 dogs (n = 1 control and n = 8 cases – elbow dysplasia, rupture of the anterior cruciate ligament and severe osteoarthritis with osteophytosis) and 6 pigs (control n = 6). Cytologic smears of synovial fluid, which proved accurate diagnoses for possible inflammatory and degenerative changes, were performed. The study of the peptide profile through the Profiling method led to differentiate between species by detecting specific peptides: in horses 1516 and 2322 Da; in dogs 2846 and 2156 Da; and finally in pigs 1308 and 2532 Da. Horses belonging to the control group did not show differences in the peptide profile between joints. The comparative peptide study in horses, including control and case groups, revealed the presence of a systematic peptide of 1415 Da in different pathological joints, except for the controls. Regarding the dog group, the spectrometric analysis revealed a frequent appearance of an 1824 Da peptide in those animals suffering from arthritis and osteoarthritis. The group of clinically healthy pigs (same breed, sex, age and joint - knee) behaved heterogeneously. Thus, a possible 1799 Da marker of joint disease was detected in animals cytologically diagnosed with inflammatory and degenerative changes, which was absent on the cytologically healthy group. The MALDI-TOF profiling is a valid, rapid and sensitive method for the analysis of peptide profile in synovial fluid of different domestic species

Álvaro Fernández Manzano

TITLE: Método de Profiling (Maldi Tof) en el líquido sinovial de caballo, perro y cerdo: Puesta a punto y estudio del perfil peptídico con ZipTip® C18

TYPE: Master Thesis

AUTHOR: Álvaro Fernández Manzano

DIRECTORS: Tellez S. and Rodriguez-Bertos A.

DATE: September 17th, 2014



Álvaro Fernández Manzano. Método de Profiling (Maldi Tof) en el líquido sinovial de caballo, perro y cerdo: Puesta a punto y estudio del perfil peptídico con ZipTip® C18 . Universidad Complutense de Madrid. September 17th, 2014. (Master Thesis)