The apparent molecular mass of samples was estimated by fitting the retention/elution volume to the calibration curve obtained from the molecular mass standards.
The average molecular masses for samples of different origin were found to be between 36 and 69 MDa, and the radius of gyration was between 130 and 170 nm.
As also shown in Figure 2, this antiserum reacted with a protein of the expected molecular mass in representative samples of immunoaffinity-purified synovial fluid from patients with psoriatic arthritis, reactive arthritis, gout, and ankylosing spondylitis.
The molecular mass of the sample was measured by MALDI-TOF (Voyager DE-STR, Applied Biosystems, Poster, CA, USA) in a linear mode with delayed extraction (75 laser shots) and an acceleration voltage of 65 kV.
The theoretical [31] density (D x ) values were calculated using the formula (4), D x = Z × M / N × a 2 × c g cm - 3 (4)where, 'Z' is the number of chemical species in the unit cell, 'M' is the molecular mass of the sample (g/mol), 'N' is the Avogadro's number (6.022 × 1023) and 'a' and 'c' are the lattice constants (cm).
In this case, the mass-averaged molecular mass of a sample with an average elution peak concentration of 2.5 μM is 45000±0.8% Da, i.e. only marginally bigger than the value expected for the monomer (42000 Da).
The raw data (in millidegrees) were converted to mean residue ellipticity (MRE), expressed in θ degrees square centimeter per decimole using the following equation 1where θ is the observed ellipticity in millidegrees, M is the molecular mass of the sample, C is the concentration in milligrams per milliliter, l is the optical path length in centimeters, and n is the number of residues.
Therefore, to determine some structural differences related with the molecular mass of Chitosan, the samples CA-V-1A and CA-V-1B were analyzed by solid state 13C-NMR spectroscopy with an 11.7 Tesla Bruker Avance III equipment.
An immunoreactive protein migrating at the expected molecular mass was detected in samples prepared from M. smegmatis containing pTZ1066 (Figure 2A) but not pSE100 (data not shown).
Using tetherin-specific polyclonal antisera, a series of bands representing variably-glycosylated tetherin monomers were noted of 30 34 Kd molecular mass, and in some samples a slower migrating series of bands were noted (Fig. 1A).
