Native and Hydrophobically Modified Human Immunoglobulin G at the Air/Water Interface.

The adsorption of human immunoglobulin G (IgG) at the air/water interface was monitored both by the in situ radiotracer technique using [(14)C] labeled IgG and by surface tension measurements. The results reveal that adsorption of IgG from single protein systems displays bimodality due to molecular rearrangements at the interface. Above the threshold value of 1.5x10(-2) mg/ml solution concentration, adsorbed IgG molecules reoriented from the side-on to the end-on configuration. The existence of a lag time which did not appear in Gamma=f(t) curves, was observed in Pi=f(t) relationships at low protein concentrations and was due to the limits of the surface pressure technique to detect protein adsorption. The adsorption of native IgG was also carried out in the presence of a hydrophobized IgG obtained by grafting capryloyl residues to its lysine groups by reaction with N-hydroxysuccinimide ester of caprylic acid, which yielded 19 covalently bound alkyl chains to the IgG molecule (19C(8)-IgG). This modified IgG exhibited enhanced adsorption at the air/water interface, as manifested by its increased adsorption efficiency relative to the native protein. Sequential and competitive adsorption experiments from binary mixtures of native IgG and 19C(8)-IgG clearly demonstrate that the displacement of the native protein from the air/water interface strongly depended on the manner of how 19C(8)-IgG and native IgG competed with each other. When the two proteins competed simultaneously, 19C(8)-IgG predominantly occupied the available area but when native IgG was adsorbed first, for 2 h, the sequentially adsorbed 19C(8)-IgG was incapable of substantially displacing it from the interface. Copyright 2001 Academic Press.

Competitive adsorption of albumin against collagen at solution-air and solution-polyethylene interfaces.

The adsorption of human serum albumin (HSA) from the binary mixtures with collagen was monitored at solution-air and solution-polyethylene interfaces by the in situ measurements. The results clearly demonstrate that on both interfaces albumin is the only adsorbing protein within a large collagen solution concentration range. At the albumin concentration equal to 0.005 mg/mL, the presence of collagen in solution results in the enhancement of albumin adsorption at solution-air interface relative to its adsorption from the single protein system. The same phenomenon is manifested at the solution-polyethylene interface, although the increase in albumin adsorption at this interface occurs at the albumin concentration equal to 0.01 mg/mL. These results are attributed to the lowering in the solution-air and solution-polyethylene interfacial tensions, and thus to the increase in the spreading characteristics of albumin in the presence of collagen molecules. The desorption experiments carried out with a buffer solution on polyethylene surfaces reveal the irreversibility of adsorbed albumin from both the single and the binary mixtures with collagen. When after 20 h of adsorption from the solutions containing albumin only, collagen was added to these solutions or when the samples after that period of time were first rinsed with a buffer and then with a collagen solution, the amounts of albumin remaining at the surfaces were in both cases reduced by one-half.

Role of phospholipid lining on respiratory mucus clearance by cough.

Phospholipid lining, present at the respiratory mucus-mucosa interface, may have an important role in the protective function of the airways by its abhesive properties and may also facilitate mucus transport. To mimic respiratory mucus-mucosa interface, monolayers of three different forms of phosphatidylglycerol (PG) have been deposited on glass slides by the Langmuir-Blodgett technique. Mucus adhesion and clearance by cough of mucus on these PG-coated or noncoated surfaces have been analyzed and compared, using frog respiratory mucus as "normal" mucus. Among the three PG types studied, the phosphatidylglycerol distearoyl, which is the phospholipid with the longest saturated fatty acid chain, was found to significantly improve the mucus cough clearance by decreasing the mucus work of adhesion compared with the noncoated surfaces. On the other hand, phosphatidylglycerol dipalmitoyl did not improve mucus cough clearance although it decreased mucus adhesion, and phosphatidylglycerol dioleyl did not improve either mucus cough clearance or mucus adhesion.

Wettability of polymers by mucin aqueous solutions.

The wettability of poly(methyl methacrylate) and polyethylene by water and aqueous mucin solutions have been studied by sessile drop and under-water captive air bubble contact angles, respectively. From the sessile drop and octane under-water contact angles the polymer-water interfaces have been characterized in terms of works of adhesion and acid-base (polar) interactions. A large water-air contact angle hysteresis observed with poly(methyl methacrylate) surfaces has been attributed to side-chain beta relaxations of polymer ester methyl groups. The wettabilities of the polymers by mucin aqueous solutions have been studied as a function of protein concentration and related to the surface tensions. A positive slope of adhesion tension vs surface tension line, characteristic of polar surfaces, was found with poly(methyl methacrylate). By contrast, a change in the slope, explained as a change in mucin relative adsorption densities at solid/liquid and solid/vapour interfaces, was observed with polyethylene. This adhesion tension behavior appeared to be in agreement with previous data we have published concerning the quantity and state of mucin which are adsorbed to polymers characterized by different surface properties.

Collagen at interfaces. II: Competitive adsorption of collagen against albumin and fibrinogen.

Adsorption of chemically radiolabeled [14C] collagen from binary mixtures with albumin or fibrinogen was studied on the solution/air and solution/polyethylene interfaces and revealed the preferential adsorption of albumin. This phenomenon is confirmed by the data of surface tension measurements of single protein, collagen-albumin, and collagen-fibrinogen solutions. Desorption experiments clearly show that more irreversibly adsorbed collagen was found on polyethylene surfaces when adsorption was performed from collagen-fibrinogen than from collagen-albumin solutions. The combined adsorption-desorption and the surface tension data show that competitive adsorption of collagen at the hydrophobic surfaces is strongly influenced by the surface tension properties of the proteins in solution.

Collagen at interfaces. I. In situ collagen adsorption at solution/air and solution/polymer interfaces.

Collagen was isolated from rat tail tendons and acetylated with 1-14C acetic anhydride. In situ adsorption of this collagen from a buffer solution (pH = 2.7) was measured at the interfaces to air, polyethylene and polyethylene grafted with poly(maleic acid), respectively. The kinetics of adsorption were recorded for all surfaces studied and the corresponding diffusion coefficients for collagen in solution with various protein concentrations were calculated. The desorption of collagen from polymer surfaces was also studied. These experiments reveal the existence of both a reversibly and an irreversibly adsorbed collagen layer on the polymers tested. The desorption/adsorption ratio for the polyethylene is higher than that for the grafted polyethylene indicating stronger interactions of collagen with the grafted surface than with the non-modified polyethylene.

Adsorption of bovine submaxillary mucin on silicone contact lenses grafted with poly(vinyl pyrrolidone).

Bovine submaxillary mucin is considered to be an analogue of the high molecular protein present in the conjunctival mucus. This mucin was isolated from fresh salivary glands and acetylated with [1-14C]acetic anhydride. In situ adsorption of the bovine submaxillary mucin on silicone contact lenses ungrafted and grafted with poly(vinyl pyrrolidone) was performed for the first time using an original radiotracer technique. The results show that the adsorbed amounts of mucin are higher on grafted samples and that thick layers are adsorbed when mucin concentration in the bulk solution is increased. Desorption experiments reveal that in addition to the tightly adsorbed protein layer, a loosely bound mucin layer of the same thickness exists on grafted and ungrafted silicones.